Pesticidal genes and methods of use

ABSTRACT

Compositions having pesticidal activity and methods for their use are provided. Compositions include isolated and recombinant polypeptides having pesticidal activity, recombinant and synthetic nucleic acid molecules encoding the polypeptides, DNA constructs and vectors comprising the nucleic acid molecules, host cells comprising the vectors, and antibodies to the polypeptides. Nucleotide sequences encoding the polypeptides can be used in DNA constructs or expression cassettes for transformation and expression in organisms of interest. The compositions and methods provided are useful for producing organisms with enhanced pest resistance or tolerance. Transgenic plants and seeds comprising a nucleotide sequence that encodes a pesticidal protein of the invention are also provided. Such plants are resistant to insects and other pests. Methods are provided for producing the various polypeptides disclosed herein, and for using those polypeptides for controlling or killing a pest. Methods and kits for detecting polypeptides of the invention in a sample are also included.

CROSS REFERENCE TO RELATED APPLICATION

This application is a divisional application of U.S. application Ser.No. 15/158,380, filed May 18, 2016, which claims the benefit of U.S.Provisional Application Ser. No. 62/170,345, filed Jun. 3, 2015, thecontents of both of which are herein incorporated by reference in theirentirety.

FIELD OF THE INVENTION

The invention is drawn to methods and compositions for controllingpests, particularly plant pests.

REFERENCE TO A SEQUENCE LISTING SUBMITTED AS A TEXT FILE VIA EFS-WEB

The official copy of the sequence listing is submitted electronicallyvia EFS-Web as an ASCII formatted sequence listing with a file namedAgB013USDIV1_SequenceListing.txt, created on Jan. 19, 2018, and having asize of 1.14 MB and is filed concurrently with the specification. Thesequence listing contained in this ASCII formatted document is part ofthe specification and is herein incorporated by reference in itsentirety.

BACKGROUND

Pests, plant diseases, and weeds can be serious threats to crops. Lossesdue to pests and diseases have been estimated at 37% of the agriculturalproduction worldwide, with 13% due to insects, bacteria and otherorganisms.

Toxins are virulence determinants that play an important role inmicrobial pathogenicity and/or evasion of the host immune response.Toxins from the gram-positive bacterium Bacillus, particularly Bacillusthuringiensis, have been used as insecticidal proteins. Currentstrategies use the genes expressing these toxins to produce transgeniccrops. Transgenic crops expressing insecticidal protein toxins are usedto combat crop damage from insects.

While the use of Bacillus toxins has been successful in controllinginsects, resistance to Bt toxins has developed in some target pests inmany parts of the world where such toxins have been used intensively.One way of solving this problem is sowing Bt crops with alternating rowsof regular non Bt crops (refuge). An alternative method to avoid or slowdown development of insect resistance is stacking insecticidal geneswith different modes of action against insects in transgenic plants. Thecurrent strategy of using transgenic crops expressing insecticidalprotein toxins is placing increasing emphasis on the discovery of noveltoxins, beyond those already derived from the bacterium Bacillusthuringiensis. These toxins may prove useful as alternatives to thosederived from B. thuringiensis for deployment in insect- andpest-resistant transgenic plants. Thus, new toxin proteins are needed.

SUMMARY

Compositions having pesticidal activity and methods for their use areprovided. Compositions include isolated and recombinant polypeptidesequences having pesticidal activity, recombinant and synthetic nucleicacid molecules encoding the pesticidal polypeptides, DNA constructscomprising the nucleic acid molecules, vectors comprising the nucleicacid molecules, host cells comprising the vectors, and antibodies to thepesticidal polypeptides. Nucleotide sequences encoding the polypeptidesprovided herein can be used in DNA constructs or expression cassettesfor transformation and expression in organisms of interest, includingmicroorganisms and plants.

The compositions and methods provided herein are useful for theproduction of organisms with enhanced pest resistance or tolerance.These organisms and compositions comprising the organisms are desirablefor agricultural purposes. Transgenic plants and seeds comprising anucleotide sequence that encodes a pesticidal protein of the inventionare also provided. Such plants are resistant to insects and other pests.

Methods are provided for producing the various polypeptides disclosedherein, and for using those polypeptides for controlling or killing apest. Methods and kits for detecting polypeptides of the invention in asample are also included.

DETAILED DESCRIPTION OF THE INVENTION

The present inventions now will be described more fully hereinafter.Indeed, these inventions may be embodied in many different forms andshould not be construed as limited to the embodiments set forth herein;rather, these embodiments are provided so that this disclosure willsatisfy applicable legal requirements.

Many modifications and other embodiments of the inventions set forthherein will come to mind to one skilled in the art to which theseinventions pertain having the benefit of the teachings presented in theforegoing descriptions. Therefore, it is to be understood that theinventions are not to be limited to the specific embodiments disclosedand that modifications and other embodiments are intended to be includedwithin the scope of the appended claims. Although specific terms areemployed herein, they are used in a generic and descriptive sense onlyand not for purposes of limitation.

I. Polynucleotides and Polypeptides

Compositions and method for conferring pesticidal activity to anorganism are is provided. The modified organism exhibits pesticidalresistance or tolerance. Recombinant pesticidal proteins, orpolypeptides and fragments and variants thereof that retain pesticidalactivity, are provided and include those set forth in SEQ ID NOs: 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40,41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58,59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76,77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109,110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123,124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137,138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151,152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165,166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179,180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193,194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207,208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221,222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235,236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249,250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263,264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277,278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291,292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305,306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319,320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333,334, 335, 336, 337, 338, 339, 340, and/or 341. The pesticidal proteinsare biologically active (e.g., pesticidal) against pests includinginsects, fungi, nematodes, and the like. Nucleotides encoding thepesticidal polypeptides, including for example, SEQ ID NOS: 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59,60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77,78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110,111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124,125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138,139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152,153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166,167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180,181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194,195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208,209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222,223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236,237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250,251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264,265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278,279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292,293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306,307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320,321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334,335, 336, 337, 338, 339, 340, and/or 341 or active fragments or variantsthereof, can be used to produce transgenic organisms, such as plants andmicroorganisms. The pesticidal proteins are biologically active (forexample, are pesticidal) against pests including insects, fungi,nematodes, and the like. Polynucleotides encoding the pesticidalpolypeptides, including for example, SEQ ID NOS: 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44,45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62,63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98,99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112,113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126,127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140,141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154,155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168,169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196,197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210,211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224,225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238,239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252,253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266,267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280,281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294,295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308,309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322,323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336,337, 338, 339, 340, and/or 341 or active fragments or variants thereof,can be used to produce transgenic organisms, such as plants andmicroorganisms. The transformed organisms are characterized by genomesthat comprise at least one stably incorporated DNA construct comprisinga coding sequence for a pesticidal protein disclosed herein. In someembodiments, the coding sequence is operably linked to a promoter thatdrives expression of the encoded pesticidal polypeptide. Accordingly,transformed microorganisms, plant cells, plant tissues, plants, seeds,and plant parts are provided. A summary of various polypeptides, activevariants and fragments thereof, and polynucleotides encoding the sameare set forth below in Table 1. As noted in Table 1, various forms ofpolypeptides are provided. Full length pesticidal polypeptides, as wellas, modified versions of the original full-length sequence (i.e.,variants) are provided. Table 1 further denotes “CryBP1” sequences. Suchsequences comprise accessory polypeptides that can be associated withsome of the toxin genes. In such instances, the CryBP1 sequences can beused alone or in combination with any of the pesticidal polypeptidesprovided herein. Table 1 further provides Split-Cry C-terminuspolypeptides. Such sequences comprise the sequence of a downstreamprotein that has homology to the C-terminal end of the Cry class oftoxin genes and are usually found after a Cry gene that is notfull-length and is missing the expected C-terminal region.

TABLE 1 Summary of SEQ ID NOs, Gene Class, and Variants thereofPolypeptides of the invention Split-Cry (and polynucleotides encodingPolypeptides of the invention Full-length Modified CryBP1 C-terminus thesame) include those having (and polynucleotides encoding SEQ ID SEQIDSEQ ID SEQ ID the % sequence identity listed the same) include thosehaving Gene Name No. No.(s) No. No. Homologs Gene Class below thesimilarity set forth below APG00038 1 APG00181 (90.3% identity, 94.6%similarity) Vip 30, 35, 40, 45, 50, 55, 60, 65, 70, 45, 50, 55, 60, 65,70, 75, 80, 75, 80, 85, 90, 95, 96, 97, 98, 99 85, 90, 95, 96, 97, 98,99 APG00131 (60.1% identity, 75.4% similarity) Vip3Ca1 (28.9% identity,42.7% similarity) APG00138 2 204 APG00108 (71.8% identity, 72.4%similarity) Cyt 60, 65, 70, 75, 80, 85, 90, 95, 96, 60, 65, 70, 75, 80,85, 90, 95, 97, 98, 99 96, 97, 98, 99 WP_030313032.1 (58.5% identity,59.1% similarity) WP_015038174.1 (57.2% identity, 58.2% similarity)APG00134 (56.6% identity, 57.2% similarity) WP_030212110.1 (55.1%identity, 56.9% similarity) APG00198 (51.1% identity, 54.5% similarity)Cyt2Ba10 (16.5% identity, 30.0% similarity) APG00139 3 WP_030030269.1(77.6% identity, 86.3% similarity) Cry6 80, 85, 90, 95, 96, 97, 98, 9990, 95, 96, 97, 98, 99 WP_001224941.1 (75.6% identity, 84.3% similarity)WP_023524015.1 (75.3% identity, 84.0% similarity) US_8802420_B2-45_1(31.6% identity, 49.1% similarity) APG00143 4 205 APG00441 (88.4%identity, 92.8% similarity) Mtx 35, 40, 45, 50, 55, 60, 65, 70, 75, 55,60, 65, 70, 75, 80, 85, 90, 80, 85, 90, 95, 96, 97, 98, 99 95, 96, 97,98, 99 APG00444 (87.8% identity, 92.5% similarity) APG00833 (80.4%identity, 87.6% similarity) WP_006918908.1 (32.0% identity, 52.6%similarity) APG00154 5 206 APG00594 (98.0% identity, 98.6% similarity)Mtx 55, 60, 65, 70, 75, 80, 85, 90, 95, 70, 75, 80, 85, 90, 95, 96, 97,96, 97, 98, 99 98, 99 APG00486 (76.5% identity, 87.3% similarity)APG00421 (63.0% identity, 74.9% similarity) APG00483 (62.6% identity,75.1% similarity) US20130227743A1_110 (51.8% identity, 68.1% similarity)AGP17985.1 (46.4% identity, 60.9% similarity) AGP17983.1 (45.1%identity, 61.9% similarity) WP_002166885.1 (39.7% identity, 56.9%similarity) APG00156 6 207 US20130227743A1_114 (27.3% identity, 44.7%Mtx 30, 35, 40, 45, 50, 55, 60, 65, 70, 45, 50, 55, 60, 65, 70, 75, 80,similarity) 75, 80, 85, 90, 95, 96, 97, 98, 99 85, 90, 95, 96, 97, 98,99 US20130227743A1_112 (27.0% identity, 43.8% similarity) AIK29697.1(25.0% identity, 41.2% similarity) WP_000239374.1 (24.9% identity, 39.7%similarity) APG00157 7 208 APG00356 (97.9% identity, 98.9% similarity)Bin 85, 90, 95, 96, 97, 98, 99 95, 96, 97, 98, 99 APG00287 (97.1%identity, 98.1% similarity) APG00377 (95.2% identity, 97.9% similarity)APG00231 (93.6% identity, 97.1% similarity) APG00035 (89.0% identity,93.6% similarity) APG00669 (86.6% identity, 93.3% similarity)WP_000143308.1 (84.7% identity, 90.9% similarity) WP_000143307.1 (84.5%identity, 91.2% similarity) US20130227743A1_6 (79.1% identity, 84.2%similarity) APG00257 (50.5% identity, 69.9% similarity) Cry35Ab4 (23.3%identity, 39.6% similarity) APG00158 8 209 APG00426 (63.8% identity,72.4% similarity) Mtx 50, 55, 60, 65, 70, 75, 80, 85, 90, 60, 65, 70,75, 80, 85, 90, 95, 95, 96, 97, 98, 99 96, 97, 98, 99 APG00517 (62.3%identity, 69.5% similarity) APG00891 (61.8% identity, 69.3% similarity)APG00227 (59.8% identity, 67.4% similarity) APG00704 (52.4% identity,65.5% similarity) APG00020 (50.1% identity, 62.4% similarity)WP_002166885.1 (46.1% identity, 57.3% similarity) US20130227743A1_110(39.3% identity, 55.3% similarity) AGP17985.1 (33.9% identity, 50.4%similarity) AGP17983.1 (29.4% identity, 45.5% similarity) APG00159 9WP_000022219.1 (77.0% identity, 87.0% similarity) Mtx 80, 85, 90, 95,96, 97, 98, 99 90, 95, 96, 97, 98, 99 AGP18072.1 (76.0% identity, 85.9%similarity) Cry64Aa1 (32.5% identity, 51.4% similarity) APG00160 10WP_003209431.1 (93.6% identity, 96.7% similarity) Cry6 95, 96, 97, 98,99 97, 98, 99 WP_018783608.1 (93.6% identity, 96.7% similarity)WP_003204425.1 (93.3% identity, 96.4% similarity) WP_033798411.1 (93.3%identity, 96.4% similarity) APG00197 (86.1% identity, 92.5% similarity)APG00286 (52.7% identity, 73.3% similarity) APG00165 11  12, 210, 211Cry21Ca2 (56.9% identity, 65.5% similarity) Cry21 60, 65, 70, 75, 80,85, 90, 95, 96, 70, 75, 80, 85, 90, 95, 96, 97, 97, 98, 99 98, 99APG00170 13  14 APG00293 (77.2% identity, 82.6% similarity) Mtx 50, 55,60, 65, 70, 75, 80, 85, 90, 65, 70, 75, 80, 85, 90, 95, 96, 95, 96, 97,98, 99 97, 98, 99 APG00051 (50.8% identity, 64.6% similarity)US_8829279_B2-11 (48.7% identity, 61.9% similarity) AGP17983.1 (31.7%identity, 45.3% similarity) WP_000844424.1 (30.1% identity, 44.3%similarity) WP_000844425.1 (29.9% identity, 44.1% similarity) APG0017115 212, 213 US20130227743A1_122 (22.8% identity, 35.7% Mtx 25, 30, 35,40, 45, 50, 55, 60, 65, 40, 45, 50, 55, 60, 65, 70, 75, similarity) 70,75, 80, 85, 90, 95, 96, 97, 98, 80, 85, 90, 95, 96, 97, 98, 99 99WP_000715637.1 (22.2% identity, 35.7% similarity) WP_016123567.1 (20.2%identity, 34.9% similarity) US20130227743A1_118 (17.6% identity, 30.7%similarity) APG00181 16 APG00038 (90.3% identity, 94.6% similarity) Vip30, 35, 40, 45, 50, 55, 60, 65, 70, 45, 50, 55, 60, 65, 70, 75, 80, 75,80, 85, 90, 95, 96, 97, 98, 99 85, 90, 95, 96, 97, 98, 99 APG00131(59.0% identity, 75.1% similarity) Vip3Ag4 (28.7% identity, 42.5%similarity) APG00182 17  18, 214, 215 WP_017251277.1 (28.2% identity,41.7% similarity) Cry 30, 35, 40, 45, 50, 55, 60, 65, 70, 45, 50, 55,60, 65, 70, 75, 80, 75, 80, 85, 90, 95, 96, 97, 98, 99 85, 90, 95, 96,97, 98, 99 WP_016742208.1 (28.1% identity, 41.9% similarity)WP_015892933.1 (28.1% identity, 40.6% similarity) Cry5Ba1 (17.4%identity, 26.3% similarity) APG00184 19 US20130227743A1_206 (19.5%identity, 30.4% Cry 20, 25, 30, 35, 40, 45, 50, 55, 60, 35, 40, 45, 50,55, 60, 65, 70, similarity) 65, 70, 75, 80, 85, 90, 95, 96, 97, 75, 80,85, 90, 95, 96, 97, 98, 98, 99 99 AGA40057.1 (19.4% identity, 29.8%similarity) AGA40058.1 (17.5% identity, 27.0% similarity) AGP17992.1(16.8% identity, 25.5% similarity) APG00187 20  21 APG00218 (56.6%identity, 71.0% similarity) Mtx 30, 35, 40, 45, 50, 55, 60, 65, 70, 45,50, 55, 60, 65, 70, 75, 80, 75, 80, 85, 90, 95, 96, 97, 98, 99 85, 90,95, 96, 97, 98, 99 APG00245 (56.3% identity, 70.4% similarity)WP_011254649.1 (27.5% identity, 44.5% similarity) AGA40032.1 (20.2%identity, 36.1% similarity) J8KZ77_BACCE (19.6% identity, 35.5%similarity) APG00193 22 WP_035088343.1 (32.7% identity, 46.4%similarity) Bin 35, 40, 45, 50, 55, 60, 65, 70, 75, 50, 55, 60, 65, 70,75, 80, 85, 80, 85, 90, 95, 96, 97, 98, 99 90, 95, 96, 97, 98, 99WP_035074455.1 (32.6% identity, 46.2% similarity) WP_026555044.1 (31.6%identity, 46.4% similarity) WP_028278541.1 (29.7% identity, 43.4%similarity) APG00197 23 KFN02705.1 (90.2% identity, 94.9% similarity)Cry6 95, 96, 97, 98, 99 95, 96, 97, 98, 99 WP_002035072.1 (89.7%identity, 94.9% similarity) WP_003209431.1 (89.7% identity, 94.9%similarity) WP_003204425.1 (89.5% identity, 94.6% similarity) APG00160(86.1% identity, 92.5% similarity) APG00286 (52.4% identity, 73.0%similarity) APG00217 24  25 Cry5Ba1 (53.5% identity, 63.7% similarity)Cry5 55, 60, 65, 70, 75, 80, 85, 90, 95, 65, 70, 75, 80, 85, 90, 95, 96,96, 97, 98, 99 97, 98, 99 APG00218 26 216 APG00245 (97.6% identity,98.8% similarity) Mtx 35, 40, 45, 50, 55, 60, 65, 70, 75, 55, 60, 65,70, 75, 80, 85, 90, 80, 85, 90, 95, 96, 97, 98, 99 95, 96, 97, 98, 99APG00187 (56.6% identity, 71.0% similarity) WP_011254649.1 (32.8%identity, 51.0% similarity) AGA40032.1 (22.3% identity, 36.2%similarity) H3SAM9_9BACL (22.2% identity, 37.6% similarity)WP_002193629.1 (21.6% identity, 36.2% similarity) APG00220 27 217APG00428 (83.3% identity, 88.8% similarity) Mtx 45, 50, 55, 60, 65, 70,75, 80, 85, 60, 65, 70, 75, 80, 85, 90, 95, 90, 95, 96, 97, 98, 99 96,97, 98, 99 AGP17985.1 (42.9% identity, 57.6% similarity) AGP17983.1(40.9% identity, 56.6% similarity) US20130227743A1_110 (34.2% identity,50.1% similarity) US_8829279_B2-11 (31.8% identity, 47.4% similarity)APG00224 28 US_8829279_B2-37 (63.2% identity, 76.6% Mtx 65, 70, 75, 80,85, 90, 95, 96, 97, 80, 85, 90, 95, 96, 97, 98, 99 similarity) 98, 99AGA40030.1 (62.0% identity, 75.4% similarity) APG00846 (52.8% identity,70.9% similarity) US_8829279_B2-27 (49.7% identity, 67.3% similarity)CAA67205.1 (43.9% identity, 60.3% similarity) APG00225 29 218WP_003209429.1 (76.5% identity, 83.8% similarity) Cry6 80, 85, 90, 95,96, 97, 98, 99 85, 90, 95, 96, 97, 98, 99 WP_003204428.1 (76.3%identity, 83.8% similarity) WP_018783610.1 (76.3% identity, 82.7%similarity) WP_018767551.1 (76.0% identity, 83.5% similarity) APG0022630 APG00349 (97.8% identity, 99.2% similarity) Bin 45, 50, 55, 60, 65,70, 75, 80, 85, 65, 70, 75, 80, 85, 90, 95, 96, 90, 95, 96, 97, 98, 9997, 98, 99 APG00797 (90.7% identity, 95.1% similarity) APG00257 (57.2%identity, 73.2% similarity) WP_000143308.1 (43.4% identity, 61.9%similarity) WP_000143307.1 (42.7% identity, 61.8% similarity)US20130227743A1_6 (40.4% identity, 59.6% similarity) Cry35Ac1 (27.2%identity, 43.6% similarity) APG00228 31 219, 220 AGP17984.1 (38.5%identity, 50.7% similarity) Mtx 40, 45, 50, 55, 60, 65, 70, 75, 80, 60,65, 70, 75, 80, 85, 90, 95, 85, 90, 95, 96, 97, 98, 99 96, 97, 98, 99AGS78124.1 (37.5% identity, 55.0% similarity) AGP18033.1 (35.9%identity, 50.0% similarity) US_8829279_B2-11 (31.9% identity, 47.3%similarity) APG00236 32 AGP18071.1 (21.7% identity, 34.6% similarity)Mtx 25, 30, 35, 40, 45, 50, 55, 60, 65, 35, 40, 45, 50, 55, 60, 65, 70,70, 75, 80, 85, 90, 95, 96, 97, 98, 75, 80, 85, 90, 95, 96, 97, 98, 9999 APG00244 33 221 APG00541 (94.6% identity, 97.4% similarity) Mtx 55,60, 65, 70, 75, 80, 85, 90, 95, 75, 80, 85, 90, 95, 96, 97, 98, 96, 97,98, 99 99 APG00509 (93.3% identity, 96.8% similarity) APG00896 (58.0%identity, 73.7% similarity) WP_016078640.1 (53.4% identity, 72.8%similarity) WP_036654376.1 (47.5% identity, 62.6% similarity) ETK27180.1(46.5% identity, 61.5% similarity) WP_033699741.1 (45.6% identity, 62.6%similarity) APG00245 34 222 APG00218 (97.6% identity, 98.8% similarity)Mtx 35, 40, 45, 50, 55, 60, 65, 70, 75, 55, 60, 65, 70, 75, 80, 85, 90,80, 85, 90, 95, 96, 97, 98, 99 95, 96, 97, 98, 99 APG00187 (56.3%identity, 70.4% similarity) WP_011254649.1 (32.0% identity, 50.0%similarity) H3SAM9_9BACL (21.6% identity, 37.6% similarity) M9M4X1_PAEPP(21.3% identity, 38.4% similarity) WP_002193629.1 (19.7% identity, 33.7%similarity) APG00246 35 223, 224 AGP17985.1 (54.8% identity, 69.4%similarity) Mtx 55, 60, 65, 70, 75, 80, 85, 90, 95, 70, 75, 80, 85, 90,95, 96, 97, 96, 97, 98, 99 98, 99 AGP17983.1 (49.9% identity, 67.0%similarity) US20130227743A1_110 (39.4% identity, 56.4% similarity)WP_002166885.1 (33.2% identity, 49.9% similarity) APG00247 36 225APG00248 (96.4% identity, 98.6% similarity) Bin 65, 70, 75, 80, 85, 90,95, 96, 97, 75, 80, 85, 90, 95, 96, 97, 98, 98, 99 99 APG00262 (63.6%identity, 73.7% similarity) WP_002191947.1 (60.2% identity, 70.1%similarity) WP_002166959.1 (60.0% identity, 70.1% similarity) AGA40039.1(59.2% identity, 70.0% similarity) APG00243 (58.6% identity, 69.2%similarity) APG00369 (57.8% identity, 68.6% similarity) APG00065 (55.8%identity, 67.4% similarity) APG00300 (53.1% identity, 64.8% similarity)Cry36Aa1 (20.3% identity, 33.4% similarity) APG00250 37 APG00053 (99.0%identity, 99.4% similarity) Cry 30, 35, 40, 45, 50, 55, 60, 65, 70, 45,50, 55, 60, 65, 70, 75, 80, 75, 80, 85, 90, 95, 96, 97, 98, 99 85, 90,95, 96, 97, 98, 99 WP_017762616.1 (28.2% identity, 44.0% similarity)AGA40058.1 (25.4% identity, 42.6% similarity) US20130227743A1_200 (22.9%identity, 38.9% similarity) Cry19Ca1 (9.6% identity, 17.1% similarity)APG00252 38 226 APG00277 (65.4% identity, 80.1% similarity) Mtx 60, 65,70, 75, 80, 85, 90, 95, 96, 75, 80, 85, 90, 95, 96, 97, 98, 97, 98, 9999 APG00006 (58.0% identity, 75.6% similarity) APG00067 (57.3% identity,76.2% similarity) APG00036 (56.8% identity, 76.6% similarity) APG00107(56.1% identity, 74.4% similarity) US20130227743A1_100 (56.1% identity,72.8% similarity) WP_000963933.1 (55.1% identity, 72.8% similarity)APG00260 (54.6% identity, 74.8% similarity) APG00201 (52.9% identity,73.9% similarity) US20130227743A1_60 (34.5% identity, 44.8% similarity)AGA40045.1 (24.5% identity, 39.0% similarity) APG00253 39 227 APG00418(95.8% identity, 96.4% similarity) Mtx 80, 85, 90, 95, 96, 97, 98, 9985, 90, 95, 96, 97, 98, 99 APG00020 (89.9% identity, 94.1% similarity)APG00764 (83.8% identity, 88.0% similarity) APG00481 (81.4% identity,87.0% similarity) APG00268 (80.9% identity, 86.4% similarity) APG00635(80.3% identity, 84.6% similarity) WP_002166885.1 (78.2% identity, 82.7%similarity) APG00227 (55.0% identity, 65.1% similarity)US20130227743A1_110 (46.2% identity, 64.2% similarity) AGP17985.1 (37.1%identity, 52.7% similarity) AGP17983.1 (35.8% identity, 53.4%similarity) APG00254 40 228 W2EAY8_9BACL (48.1% identity, 66.4%similarity) Mtx 50, 55, 60, 65, 70, 75, 80, 85, 90, 70, 75, 80, 85, 90,95, 96, 97, 95, 96, 97, 98, 99 98, 99 WP_036657032.1 (43.7% identity,59.7% similarity) W2EAU2_9BACL (38.3% identity, 52.2% similarity)ETK27180.1 (33.8% identity, 45.4% similarity) APG00256 41 229 AGP18021.1(65.3% identity, 79.0% similarity) Bin 70, 75, 80, 85, 90, 95, 96, 97,98, 80, 85, 90, 95, 96, 97, 98, 99 99 AGP18019.1 (30.1% identity, 47.0%similarity) US_8461421_B2-5_1 (29.8% identity, 47.0% similarity)Cry35Ab4 (24.5% identity, 38.5% similarity) APG00257 42 APG00226 (57.2%identity, 73.2% similarity) Bin 55, 60, 65, 70, 75, 80, 85, 90, 95, 75,80, 85, 90, 95, 96, 97, 98, 96, 97, 98, 99 99 APG00349 (57.2% identity,72.4% similarity) APG00797 (56.9% identity, 73.4% similarity)WP_000143307.1 (53.0% identity, 70.8% similarity) WP_000143308.1 (52.7%identity, 70.7% similarity) APG00731 (51.6% identity, 68.8% similarity)APG00231 (51.1% identity, 69.7% similarity) APG00669 (51.1% identity,69.1% similarity) APG00377 (50.8% identity, 69.7% similarity) APG00157(50.5% identity, 69.9% similarity) US20130227743A1_6 (46.8% identity,65.4% similarity) Cry35Ac1 (24.6% identity, 43.0% similarity) APG0025943 230 WP_002140073.1 (94.6% identity, 97.7% similarity) Cry6 95, 96,97, 98, 99 98, 99 WP_016083929.1 (94.0% identity, 97.4% similarity)WP_033674722.1 (60.9% identity, 78.2% similarity) WP_018765384.1 (59.8%identity, 76.9% similarity) APG00298 (56.2% identity, 74.6% similarity)APG00260 44 231 APG00201 (90.8% identity, 94.4% similarity) Mtx 75, 80,85, 90, 95, 96, 97, 98, 99 85, 90, 95, 96, 97, 98, 99 APG00006 (78.7%identity, 85.6% similarity) APG00022 (77.5% identity, 86.3% similarity)APG00036 (75.7% identity, 83.0% similarity) APG00067 (73.4% identity,83.3% similarity) US20130227743A1_100 (73.0% identity, 82.2% similarity)WP_000963933.1 (72.5% identity, 82.3% similarity) APG00277 (61.8%identity, 76.0% similarity) APG00252 (54.6% identity, 74.8% similarity)US20130227743A1_60 (45.6% identity, 50.8% similarity) A8LVM9_SALAI(23.9% identity, 37.5% similarity) APG00268 45 232, 233 APG00481 (94.2%identity, 96.0% similarity) Mtx 80, 85, 90, 95, 96, 97, 98, 99 85, 90,95, 96, 97, 98, 99 APG00646 (93.4% identity, 96.5% similarity APG00322(92.8% identity, 94.4% similarity) APG00393 (90.2% identity, 93.9%similarity) APG00764 (88.3% identity, 92.3% similarity) APG00635 (87.2%identity, 90.2% similarity) APG00020 (81.6% identity, 86.7% similarity)WP_002166885.1 (77.9% identity, 83.5% similarity) APG00227 (55.4%identity, 67.1% similarity) US20130227743A1_110 (43.0% identity, 60.9%similarity) AGP17985.1 (36.9% identity, 53.8% similarity) AGP17983.1(35.7% identity, 53.0% similarity) APG00269 46 234, 235 APG00560 (96.2%identity, 96.8% similarity) Mtx 55, 60, 65, 70, 75, 80, 85, 90, 95, 70,75, 80, 85, 90, 95, 96, 97, 96, 97, 98, 99 98, 99 APG00995 (84.7%identity, 89.5% similarity) APG00414 (83.6% identity, 89.0% similarity)APG00593 (83.6% identity, 88.7% similarity) US20130227743A1_110 (52.3%identity, 67.4% similarity) AGP17985.1 (46.3% identity, 58.9%similarity) WP_002166885.1 (44.4% identity, 58.9% similarity) AGP17983.1(43.4% identity, 59.7% similarity) APG00276 47 236 APG00545 (74.9%identity, 84.7% similarity) Mtx 35, 40, 45, 50, 55, 60, 65, 70, 75, 50,55, 60, 65, 70, 75, 80, 85, 80, 85, 90, 95, 96, 97, 98, 99 90, 95, 96,97, 98, 99 APG00671 (56.6% identity, 75.2% similarity) WP_036654376.1(33.7% identity, 46.9% similarity) AGP18071.1 (33.1% identity, 49.2%similarity) WP_000794513.1 (32.8% identity, 48.9% similarity) ETK27180.1(32.6% identity, 46.2% similarity) APG00277 48 APG00252 (65.4% identity,80.1% similarity) Mtx 65, 70, 75, 80, 85, 90, 95, 96, 97, 80, 85, 90,95, 96, 97, 98, 99 98, 99 APG00036 (63.5% identity, 76.1% similarity)WP_000963933.1 (63.1% identity, 77.5% similarity) APG00137 (62.7%identity, 77.5% similarity) US20130227743A1_100 (62.4% identity, 77.2%similarity) APG00201 (62.2% identity, 76.3% similarity) APG00006 (62.0%identity, 77.0% similarity) APG00260 (61.8% identity, 76.0% similarity)APG00067 (61.3% identity, 78.3% similarity) US20130227743A1_60 (36.6%identity, 44.8% similarity) WP_026272455.1 (23.4% identity, 38.8%similarity) APG00281 49 237 APG00009 (52.6% identity, 67.1% similarity)Mtx 35, 40, 45, 50, 55, 60, 65, 70, 75, 55, 60, 65, 70, 75, 80, 85, 90,80, 85, 90, 95, 96, 97, 98, 99 95, 96, 97, 98, 99 US_8829279_B2-11(34.9% identity, 52.0% similarity) AGP17984.1 (30.6% identity, 44.0%similarity) AGP18033.1 (29.9% identity, 43.2% similarity) AGP17985.1(27.1% identity, 41.7% similarity) APG00283 50 238, 239 APG00479 (72.4%identity, 79.9% similarity) Mtx 40, 45, 50, 55, 60, 65, 70, 75, 80, 60,65, 70, 75, 80, 85, 90, 95, 85, 90, 95, 96, 97, 98, 99 96, 97, 98, 99APG00411 (62.4% identity, 73.3% similarity) APG00966 (57.9% identity,72.6% similarity) AGS78124.1 (37.0% identity, 55.1% similarity)AGP17984.1 (32.4% identity, 46.1% similarity) AGP18033.1 (32.1%identity, 49.5% similarity) WP_000844425.1 (29.0% identity, 45.5%similarity) APG00293 51 240 APG00170 (77.2% identity, 82.6% similarity)Mtx 55, 60, 65, 70, 75, 80, 85, 90, 95, 65, 70, 75, 80, 85, 90, 95, 96,96, 97, 98, 99 97, 98, 99 APG00911 (51.1% identity, 65.5% similarity)APG00854 (50.9% identity, 64.0% similarity) US_8829279_B2-11 (50.5%identity, 63.4% similarity) WP_000844424.1 (31.3% identity, 48.3%similarity) US_8461415_B2-38 (31.2% identity, 44.4% similarity)WP_000844425.1 (31.1% identity, 48.0% similarity) APG00296 52 241APG00553 (50.3% identity, 65.1% similarity) Cry 25, 30, 35, 40, 45, 50,55, 60, 65, 40, 45, 50, 55, 60, 65, 70, 75, 70, 75, 80, 85, 90, 95, 96,97, 98, 80, 85, 90, 95, 96, 97, 98, 99 99 US20130227743A1_200 (23.7%identity, 36.4% similarity) AGA40057.1 (19.1% identity, 31.7%similarity) WP_017762619.1 (18.2% identity, 31.2% similarity) AGP17992.1(18.2% identity, 28.4% similarity) APG00300 53 242 APG00065 (85.0%identity, 90.2% similarity) Bin 70, 75, 80, 85, 90, 95, 96, 97, 98, 80,85, 90, 95, 96, 97, 98, 99 99 APG00419 (84.4% identity, 92.0%similarity) APG00229 (78.7% identity, 86.8% similarity) APG00243 (78.6%identity, 85.9% similarity) APG00213 (78.6% identity, 85.1% similarity)WP_002166959.1 (68.8% identity, 79.7% similarity) WP_002191947.1 (68.6%identity, 79.7% similarity) WP_000839920.1 (68.6% identity, 79.2%similarity) APG00247 (53.1% identity, 64.8% similarity) Cry35Ac2 (22.3%identity, 35.0% similarity) APG00303 54 243, 244 AGP18033.1 (34.2%identity, 52.3% similarity) Mtx 35, 40, 45, 50, 55, 60, 65, 70, 75, 55,60, 65, 70, 75, 80, 85, 90, 80, 85, 90, 95, 96, 97, 98, 99 95, 96, 97,98, 99 AGP17984.1 (33.4% identity, 49.5% similarity) AGS78124.1 (29.5%identity, 44.1% similarity) US_8829279_B2-11 (28.5% identity, 45.2%similarity) APG00304 55 245 APG00129 (58.5% identity, 70.9% similarity)Mtx 60, 65, 70, 75, 80, 85, 90, 95, 96, 75, 80, 85, 90, 95, 96, 97, 98,97, 98, 99 99 WP_030030026.1 (57.5% identity, 70.4% similarity)US20130227743A1_106 (21.3% identity, 37.5% similarity) APG00309 56  57WP_001039209.1 (66.1% identity, 79.6% similarity) Mtx 70, 75, 80, 85,90, 95, 96, 97, 98, 80, 85, 90, 95, 96, 97, 98, 99 99 WP_018673409.1(63.0% identity, 77.8% similarity) APG00427 (62.7% identity, 76.5%similarity) APG00851 (59.0% identity, 74.7% similarity) US_8829279_B2-2(51.8% identity, 69.5% similarity) US_8829279_B2-61 (51.8% identity,69.5% similarity) APG00321 58 246 AGP17992.1 (25.9% identity, 40.8%similarity) Cry 30, 35, 40, 45, 50, 55, 60, 65, 70, 45, 50, 55, 60, 65,70, 75, 80, 75, 80, 85, 90, 95, 96, 97, 98, 99 85, 90, 95, 96, 97, 98,99 WP_017762616.1 (25.7% identity, 40.0% similarity) WP_017762581.1(25.2% identity, 39.3% similarity) US20130227743A1_200 (24.7% identity,39.0% similarity) APG00322 59 247 APG00481 (93.1% identity, 94.7%similarity) Mtx 80, 85, 90, 95, 96, 97, 98, 99 85, 90, 95, 96, 97, 98,99 APG00268 (92.8% identity, 94.4% similarity) APG00635 (90.7% identity,93.4% similarity) APG00764 (89.6% identity, 93.4% similarity) APG00959(89.3% identity, 91.7% similarity) APG00646 (89.1% identity, 92.8%similarity) APG00020 (80.3% identity, 85.6% similarity) WP_002166885.1(77.9% identity, 84.0% similarity) APG00227 (55.0% identity, 65.2%similarity) US20130227743A1_110 (43.1% identity, 61.1% similarity)AGP17985.1 (37.3% identity, 53.7% similarity) AGP17983.1 (36.3%identity, 55.4% similarity) APG00323 60 248 APG00420 (93.2% identity,96.2% similarity) Mtx 40, 45, 50, 55, 60, 65, 70, 75, 80, 60, 65, 70,75, 80, 85, 90, 95, 85, 90, 95, 96, 97, 98, 99 96, 97, 98, 99 APG00586(92.6% identity, 95.9% similarity) AGA40044.1 (35.6% identity, 55.6%similarity) WP_002203994.1 (17.5% identity, 19.3% similarity) APG0032461 249 WP_002166885.1 (44.3% identity, 59.2% similarity) Mtx 45, 50, 55,60, 65, 70, 75, 80, 85, 60, 65, 70, 75, 80, 85, 90, 95, 90, 95, 96, 97,98, 99 96, 97, 98, 99 US20130227743A1_110 (40.1% identity, 55.7%similarity) AGP17983.1 (33.7% identity, 51.0% similarity) AGP17985.1(32.6% identity, 47.9% similarity) APG00329 62 250 WP_017762581.1 (91.2%identity, 91.9% similarity) Cry 95, 96, 97, 98, 99 95, 96, 97, 98, 99AGP17992.1 (86.1% identity, 91.8% similarity) WP_017762619.1 (32.9%identity, 46.5% similarity) Cry7Aa1 (10.7% identity, 17.8% similarity)APG00330 63 251, 252 WP_034177808.1 (49.6% identity, 62.6% similarity)Cry 50, 55, 60, 65, 70, 75, 80, 85, 90, 65, 70, 75, 80, 85, 90, 95, 96,95, 96, 97, 98, 99 97, 98, 99 WP_011296566.1 (43.9% identity, 60.9%similarity) WP_039015225.1 (42.6% identity, 61.8% similarity) Cry1Ib4(18.1% identity, 27.2% similarity) APG00332 64 APG00907 (93.0% identity,96.5% similarity) Mtx 65, 70, 75, 80, 85, 90, 95, 96, 97, 80, 85, 90,95, 96, 97, 98, 99 98, 99 US_8318900_B2-191_1 (63.0% identity, 77.2%similarity) AGA40061.1 (59.6% identity, 73.7% similarity) APG00112(57.9% identity, 70.6% similarity) AGA40062.1 (54.2% identity, 69.5%similarity) Cry64Aa1 (37.5% identity, 51.1% similarity) APG00339 65 253AGA40042.1 (66.9% identity, 77.1% similarity) Mtx 70, 75, 80, 85, 90,95, 96, 97, 98, 80, 85, 90, 95, 96, 97, 98, 99 99 AGA40043.1 (61.7%identity, 71.9% similarity) WP_000844424.1 (26.8% identity, 44.1%similarity) WP_000844425.1 (26.5% identity, 42.3% similarity) APG0034166 AGP17988.1 (44.9% identity, 61.3% similarity) Mtx 45, 50, 55, 60, 65,70, 75, 80, 85, 65, 70, 75, 80, 85, 90, 95, 96, 90, 95, 96, 97, 98, 9997, 98, 99 APG00354 67 APG00704 (57.9% identity, 60.1% similarity) Mtx50, 55, 60, 65, 70, 75, 80, 85, 90, 55, 60, 65, 70, 75, 80, 85, 90, 95,96, 97, 98, 99 95, 96, 97, 98, 99 APG00020 (53.8% identity, 58.2%similarity) APG00393 (52.4% identity, 56.4% similarity) APG00268 (52.0%identity, 57.0% similarity) APG00764 (51.6% identity, 55.0% similarity)APG00635 (50.4% identity, 54.9% similarity) WP_002166885.1 (47.3%identity, 51.3% similarity) US20130227743A1_110 (30.6% identity, 43.5%similarity) AGP17985.1 (25.7% identity, 39.0% similarity) EJQ15321.1(25.2% identity, 38.2% similarity) APG00359 68  70 WP_033697745.1 (32.0%identity, 44.4% similarity) Mtx 35, 40, 45, 50, 55, 60, 65, 70, 75, 50,55, 60, 65, 70, 75, 80, 85, 80, 85, 90, 95, 96, 97, 98, 99 90, 95, 96,97, 98, 99 AGP17979.1 (31.8% identity, 46.4% similarity) CDN39471.1(30.5% identity, 43.2% similarity) Cry55Aa1 (22.3% identity, 37.1%similarity) APG00360 69 US_8318900_B2-191_1 (50.2% identity, 65.0% Mtx55, 60, 65, 70, 75, 80, 85, 90, 95, 70, 75, 80, 85, 90, 95, 96, 97,similarity) 96, 97, 98, 99 98, 99 AGA40061.1 (47.2% identity, 61.1%similarity) AGA40062.1 (44.0% identity, 60.7% similarity) Cry60Ba2(39.2% identity, 54.7% similarity) APG00367 71 254 APG00072 (54.2%identity, 64.9% similarity) Cry 30, 35, 40, 45, 50, 55, 60, 65, 70, 45,50, 55, 60, 65, 70, 75, 80, 75, 80, 85, 90, 95, 96, 97, 98, 99 85, 90,95, 96, 97, 98, 99 Cry11Aa1 (28.4% identity, 41.8% similarity)ABW89739.1 (28.4% identity, 41.8% similarity) US_6071877_A-7 (28.4%identity, 41.7% similarity) APG00371 72  73, 255 US_8829279_B2-11 (28.7%identity, 44.8% Mtx 30, 35, 40, 45, 50, 55, 60, 65, 70, 50, 55, 60, 65,70, 75, 80, 85, similarity) 75, 80, 85, 90, 95, 96, 97, 98, 99 90, 95,96, 97, 98, 99 US20130227743A1_110 (28.5% identity, 42.8% similarity)WP_000844424.1 (28.3% identity, 47.2% similarity) WP_000844425.1 (28.0%identity, 46.9% similarity) APG00382 74 256 APG00484 (87.0% identity,94.0% similarity) Mtx 50, 55, 60, 65, 70, 75, 80, 85, 90, 65, 70, 75,80, 85, 90, 95, 96, 95, 96, 97, 98, 99 97, 98, 99 WP_008180054.1 (47.2%identity, 63.0% similarity) WP_003290257.1 (45.7% identity, 59.9%similarity) WP_016099228.1 (45.4% identity, 63.2% similarity)WP_000790613.1 (45.4% identity, 60.9% similarity) APG00390 75 MtxAPG00393 76 257, 258 APG00268 (90.2% identity, 93.9% similarity) Mtx 80,85, 90, 95, 96, 97, 98, 99 85, 90, 95, 96, 97, 98, 99 APG00959 (89.0%identity, 92.5% similarity) APG00322 (88.5% identity, 92.5% similarity)APG00481 (88.0% identity, 92.6% similarity) APG00646 (87.5% identity,92.8% similarity) APG00635 (85.1% identity, 90.4% similarity) APG00020(81.4% identity, 87.2% similarity) WP_002166885.1 (79.2% identity, 84.8%similarity) APG00227 (55.3% identity, 67.1% similarity)US20130227743A1_110 (44.2% identity, 60.8% similarity) AGP17985.1 (36.7%identity, 52.8% similarity) AGP17983.1 (35.2% identity, 53.2%similarity) APG00402 77 WP_002193629.1 (54.2% identity, 75.8%similarity) Mtx 55, 60, 65, 70, 75, 80, 85, 90, 95, 80, 85, 90, 95, 96,97, 98, 99 96, 97, 98, 99 APG00755 (52.6% identity, 68.7% similarity)US20120278954A1_54 (24.8% identity, 46.0% similarity) WP_011254649.1(24.4% identity, 42.0% similarity) AGA40044.1 (21.0% identity, 37.2%similarity) APG00403 78 259, 260 AGP18019.1 (51.2% identity, 67.2%similarity) Bin 55, 60, 65, 70, 75, 80, 85, 90, 95, 70, 75, 80, 85, 90,95, 96, 97, 96, 97, 98, 99 98, 99 US_8461421_B2-5_1 (51.2% identity,67.2% similarity) US_8461421_B2-146_1 (43.2% identity, 55.6% similarity)Cry35Ac2 (23.8% identity, 42.1% similarity) APG00410 79  73WP_036065031.1 (27.8% identity, 43.6% similarity) Mtx 30, 35, 40, 45,50, 55, 60, 65, 70, 50, 55, 60, 65, 70, 75, 80, 85, 75, 80, 85, 90, 95,96, 97, 98, 99 90, 95, 96, 97, 98, 99 EPI38517.1 (27.0% identity, 47.2%similarity) US_8461421_B2-117 (25.5% identity, 40.4% similarity)AAB36657.1 (25.2% identity, 42.3% similarity) APG00411 80 261, 262APG00283 (62.4% identity, 73.3% similarity) Mtx 35, 40, 45, 50, 55, 60,65, 70, 75, 55, 60, 65, 70, 75, 80, 85, 90, 80, 85, 90, 95, 96, 97, 98,99 95, 96, 97, 98, 99 APG00479 (62.2% identity, 71.7% similarity)APG00966 (52.2% identity, 64.3% similarity) AGS78124.1 (34.9% identity,51.3% similarity) AGP18033.1 (29.5% identity, 44.5% similarity)AGP17984.1 (29.5% identity, 44.1% similarity) US_8829279_B2-11 (27.5%identity, 44.7% similarity) APG00414 81 263 APG00995 (98.1% identity,98.6% similarity) Mtx 55, 60, 65, 70, 75, 80, 85, 90, 95, 75, 80, 85,90, 95, 96, 97, 98, 96, 97, 98, 99 99 APG00593 (97.5% identity, 98.6%similarity) APG00560 (83.9% identity, 89.4% similarity) APG00269 (83.6%identity, 89.0% similarity) US20130227743A1_110 (54.8% identity, 70.7%similarity) AGP17985.1 (47.5% identity, 56.4% similarity) WP_002166885.1(45.2% identity, 59.7% similarity) AGP17983.1 (43.2% identity, 57.9%similarity) APG00416 82 264 US_8829279_B2-11 (32.5% identity, 52.2% Mtx35, 40, 45, 50, 55, 60, 65, 70, 75, 55, 60, 65, 70, 75, 80, 85, 90,similarity) 80, 85, 90, 95, 96, 97, 98, 99 95, 96, 97, 98, 99 AGP17983.1(32.0% identity, 45.7% similarity) WP_000844424.1 (30.3% identity, 48.9%similarity) WP_000844425.1 (30.3% identity, 48.9% similarity) APG0041783 265 APG00667 (62.3% identity, 73.0% similarity) Mtx 55, 60, 65, 70,75, 80, 85, 90, 95, 70, 75, 80, 85, 90, 95, 96, 97, 96, 97, 98, 99 98,99 APG00938 (53.8% identity, 69.4% similarity) APG00590 (53.7% identity,68.9% similarity) APG00563 (53.0% identity, 67.1% similarity) APG00794(52.7% identity, 67.4% similarity) WP_000794514.1 (51.0% identity, 65.8%similarity) US20130227743A1_102 (50.4% identity, 67.2% similarity)APG00146 (50.1% identity, 64.9% similarity) AGA40045.1 (47.6% identity,61.8% similarity) WP_036654376.1 (39.6% identity, 52.0% similarity)APG00418 84 266 APG00253 (95.8% identity, 96.4% similarity) Mtx 80, 85,90, 95, 96, 97, 98, 99 80, 85, 90, 95, 96, 97, 98, 99 APG00020 (87.4%identity, 91.3% similarity) APG00764 (80.6% identity, 85.1% similarity)APG00481 (78.8% identity, 84.4% similarity) APG00268 (78.5% identity,83.5% similarity) APG00635 (78.0% identity, 82.0% similarity)WP_002166885.1 (75.5% identity, 79.8% similarity) APG00227 (53.5%identity, 63.3% similarity) US20130227743A1_110 (43.2% identity, 61.5%similarity) AGP17985.1 (35.2% identity, 50.9% similarity) AGP17983.1(33.4% identity, 51.1% similarity) APG00420 85 267, 268 APG00586 (95.6%identity, 96.8% similarity) Mtx 40, 45, 50, 55, 60, 65, 70, 75, 80, 55,60, 65, 70, 75, 80, 85, 90, 85, 90, 95, 96, 97, 98, 99 95, 96, 97, 98,99 APG00323 (93.2% identity, 96.2% similarity) AGA40044.1 (36.0%identity, 54.7% similarity) WP_002203994.1 (16.5% identity, 18.8%similarity) APG00421 86 269 APG00483 (85.8% identity, 93.2% similarity)Mtx 70, 75, 80, 85, 90, 95, 96, 97, 98, 80, 85, 90, 95, 96, 97, 98, 9999 APG00995 (67.9% identity, 79.2% similarity) APG00593 (67.1% identity,79.2% similarity) APG00414 (67.0% identity, 78.9% similarity)US20130227743A1_110 (65.9% identity, 77.2% similarity) WP_002166885.1(44.8% identity, 59.3% similarity) AGP17985.1 (44.1% identity, 61.2%similarity) AGP17983.1 (43.5% identity, 59.8% similarity) APG00425 87APG00645 (71.8% identity, 80.2% similarity) Mtx 40, 45, 50, 55, 60, 65,70, 75, 80, 60, 65, 70, 75, 80, 85, 90, 95, 85, 90, 95, 96, 97, 98, 9996, 97, 98, 99 AGA40044.1 (37.7% identity, 55.7% similarity) APG00426 88270 APG00891 (93.9% identity, 94.6% similarity) Mtx 55, 60, 65, 70, 75,80, 85, 90, 95, 65, 70, 75, 80, 85, 90, 95, 96, 96, 97, 98, 99 97, 98,99 APG00227 (91.0% identity, 91.8% similarity) APG00517 (81.3% identity,83.4% similarity) APG00158 (63.8% identity, 72.4% similarity) APG00253(57.9% identity, 68.9% similarity) APG00020 (56.6% identity, 68.7%similarity) APG00481 (56.1% identity, 66.4% similarity) APG00635 (54.5%identity, 65.6% similarity) WP_002166885.1 (52.3% identity, 61.8%similarity) US20130227743A1_110 (42.7% identity, 57.5% similarity)AGP17985.1 (36.4% identity, 52.3% similarity) US_8829279_B2-11 (32.7%identity, 46.8% similarity) APG00428 89 271 APG00220 (83.3% identity,88.8% similarity) Mtx 45, 50, 55, 60, 65, 70, 75, 80, 85, 60, 65, 70,75, 80, 85, 90, 95, 90, 95, 96, 97, 98, 99 96, 97, 98, 99 AGP17985.1(41.8% identity, 57.4% similarity) AGP17983.1 (41.6% identity, 57.4%similarity) US20130227743A1_110 (33.2% identity, 49.5% similarity)US_8829279_B2-11 (31.2% identity, 47.1% similarity) APG00432 90WP_000240775.1 (73.1% identity, 87.0% similarity) Cry6 75, 80, 85, 90,95, 96, 97, 98, 99 90, 95, 96, 97, 98, 99 WP_000240776.1 (73.1%identity, 86.4% similarity) WP_002144456.1 (73.1% identity, 86.4%similarity) APG00342 (72.8% identity, 86.1% similarity) APG00024 (71.6%identity, 84.7% similarity) APG00180 (64.0% identity, 81.0% similarity)Cry6Ba1 (27.0% identity, 44.2% similarity) APG00441 91 272 APG00444(98.4% identity, 99.1% similarity) Mtx 35, 40, 45, 50, 55, 60, 65, 70,75, 55, 60, 65, 70, 75, 80, 85, 90, 80, 85, 90, 95, 96, 97, 98, 99 95,96, 97, 98, 99 APG00143 (88.4% identity, 92.8% similarity) APG00833(79.4% identity, 86.9% similarity) WP_006918908.1 (31.8% identity, 51.5%similarity) APG00443 92 273 APG00510 (72.5% identity, 72.9% similarity)Cry 55, 60, 65, 70, 75, 80, 85, 90, 95, 75, 80, 85, 90, 95, 96, 97, 98,96, 97, 98, 99 99 WP_017762621.1 (53.9% identity, 71.3% similarity)WP_017762616.1 (19.6% identity, 30.4% similarity) AGA40058.1 (18.7%identity, 29.2% similarity) WP_017762619.1 (16.8% identity, 27.7%similarity) APG00444 93 274 APG00441 (98.4% identity, 99.1% similarity)Mtx 35, 40, 45, 50, 55, 60, 65, 70, 75, 55, 60, 65, 70, 75, 80, 85, 90,80, 85, 90, 95, 96, 97, 98, 99 95, 96, 97, 98, 99 APG00143 (87.8%identity, 92.5% similarity) APG00833 (78.8% identity, 86.3% similarity)WP_006918908.1 (31.8% identity, 51.2% similarity) APG00451 94 275US_8829279_B2-11 (41.3% identity, 54.7% Mtx 45, 50, 55, 60, 65, 70, 75,80, 85, 55, 60, 65, 70, 75, 80, 85, 90, similarity) 90, 95, 96, 97, 98,99 95, 96, 97, 98, 99 WP_000844424.1 (35.0% identity, 51.7% similarity)WP_000844425.1 (34.7% identity, 51.1% similarity) WP_033690552.1 (33.1%identity, 47.8% similarity) APG00453 95 276 Cry23Aa1 (20.8% identity,36.4% similarity) Mtx 25, 30, 35, 40, 45, 50, 55, 60, 65, 40, 45, 50,55, 60, 65, 70, 75, 70, 75, 80, 85, 90, 95, 96, 97, 98, 80, 85, 90, 95,96, 97, 98, 99 99 APG00455 96  97 WP_015811789.1 (23.6% identity, 36.0%similarity) Cry 25, 30, 35, 40, 45, 50, 55, 60, 65, 40, 45, 50, 55, 60,65, 70, 75, 70, 75, 80, 85, 90, 95, 96, 97, 98, 80, 85, 90, 95, 96, 97,98, 99 99 WP_026632766.1 (21.6% identity, 32.7% similarity) CCH51877.1(21.0% identity, 32.6% similarity) Cry73Aa (15.1% identity, 23.7%similarity) APG00457 98 WP_017762616.1 (30.2% identity, 45.4%similarity) Cry 35, 40, 45, 50, 55, 60, 65, 70, 75, 50, 55, 60, 65, 70,75, 80, 85, 80, 85, 90, 95, 96, 97, 98, 99 90, 95, 96, 97, 98, 99US20130227743A1_200 (27.7% identity, 40.3% similarity) AGA40057.1 (26.2%identity, 41.0% similarity) AGA40058.1 (24.3% identity, 39.6%similarity) APG00477 99 100 WP_016098287.1 (92.8% identity, 96.2%similarity) Mtx 95, 96, 97, 98, 99 97, 98, 99 EOP91365.1 (47.1%identity, 60.9% similarity) WP_016099611.1 (40.8% identity, 58.7%similarity) US_8461421_B2-117 (29.8% identity, 44.7% similarity)APG00479 101 277 APG00283 (72.4% identity, 79.9% similarity) Mtx 40, 45,50, 55, 60, 65, 70, 75, 80, 55, 60, 65, 70, 75, 80, 85, 90, 85, 90, 95,96, 97, 98, 99 95, 96, 97, 98, 99 APG00411 (62.2% identity, 71.7%similarity) APG00966 (54.0% identity, 68.9% similarity) AGS78124.1(35.9% identity, 52.8% similarity) AGP17984.1 (30.4% identity, 44.2%similarity) WP_000844425.1 (29.4% identity, 45.4% similarity) AGP18033.1(29.4% identity, 41.4% similarity) APG00480 102 278 APG00733 (71.4%identity, 80.8% similarity) Cry 25, 30, 35, 40, 45, 50, 55, 60, 65, 40,45, 50, 55, 60, 65, 70, 75, 70, 75, 80, 85, 90, 95, 96, 97, 98, 80, 85,90, 95, 96, 97, 98, 99 99 APG00553 (69.1% identity, 84.3% similarity)WP_017762616.1 (24.3% identity, 39.4% similarity) US20130227743A1_200(21.6% identity, 36.1% similarity) AGA40057.1 (21.6% identity, 34.2%similarity) WP_017762581.1 (20.9% identity, 31.0% similarity) APG00481103 279, 280 APG00268 (94.2% identity, 96.0% similarity) Mtx 80, 85, 90,95, 96, 97, 98, 99 85, 90, 95, 96, 97, 98, 99 APG00322 (93.1% identity,94.7% similarity) APG00646 (92.3% identity, 95.8% similarity) APG00959(88.8% identity, 91.5% similarity) APG00764 (88.3% identity, 92.3%similarity) APG00635 (85.9% identity, 89.4% similarity) APG00020 (80.1%identity, 86.2% similarity) WP_002166885.1 (77.2% identity, 83.6%similarity) APG00227 (56.6% identity, 66.7% similarity)US20130227743A1_110 (43.9% identity, 61.4% similarity) AGP17985.1 (38.1%identity, 54.2% similarity) AGP17983.1 (35.9% identity, 53.6%similarity) APG00483 104 281 APG00421 (85.8% identity, 93.2% similarity)Mtx 65, 70, 75, 80, 85, 90, 95, 96, 97, 80, 85, 90, 95, 96, 97, 98, 9998, 99 APG00995 (69.5% identity, 79.2% similarity) APG00593 (68.7%identity, 79.0% similarity) APG00414 (68.2% identity, 78.7% similarity)US20130227743A1_110 (63.7% identity, 75.6% similarity) APG00020 (52.0%identity, 65.4% similarity) AGP17985.1 (48.3% identity, 60.4%similarity) WP_002166885.1 (47.1% identity, 59.1% similarity) AGP17983.1(45.4% identity, 60.1% similarity) APG00484 105 282 APG00382 (87.0%identity, 94.0% similarity) Mtx 50, 55, 60, 65, 70, 75, 80, 85, 90, 65,70, 75, 80, 85, 90, 95, 96, 95, 96, 97, 98, 99 97, 98, 99 WP_008180054.1(48.7% identity, 61.4% similarity) WP_003290257.1 (46.1% identity, 57.9%similarity) WP_016099228.1 (45.7% identity, 63.1% similarity) AGP17978.1(44.5% identity, 60.5% similarity) APG00485 106 Cry22Ba1 (78.1%identity, 83.8% similarity) Cry22B 80, 85, 90, 95, 96, 97, 98, 99 85,90, 95, 96, 97, 98, 99 APG00008 (77.9% identity, 82.9% similarity)APG00486 107 283 APG00154 (76.5% identity, 87.3% similarity) Mtx 55, 60,65, 70, 75, 80, 85, 90, 95, 70, 75, 80, 85, 90, 95, 96, 97, 96, 97, 98,99 98, 99 APG00594 (75.9% identity, 86.7% similarity) APG00421 (60.8%identity, 74.6% similarity) APG00269 (59.6% identity, 72.6% similarity)US20130227743A1_110 (51.8% identity, 68.0% similarity) AGP17985.1 (44.3%identity, 61.7% similarity) AGP17983.1 (42.9% identity, 60.0%similarity) WP_002166885.1 (42.3% identity, 56.4% similarity) APG00488108 109 US20130227743A1_106 (41.6% identity, 59.6% Mtx 45, 50, 55, 60,65, 70, 75, 80, 85, 60, 65, 70, 75, 80, 85, 90, 95, similarity) 90, 95,96, 97, 98, 99 96, 97, 98, 99 W8YCZ9_BACTU (23.3% identity, 37.0%similarity) KEZ79637.1 (19.9% identity, 30.5% similarity) APG00496 110284 APG00500 (71.5% identity, 76.8% similarity) Cry 55, 60, 65, 70, 75,80, 85, 90, 95, 65, 70, 75, 80, 85, 90, 95, 96, 96, 97, 98, 99 97, 98,99 APG00113 (56.6% identity, 66.8% similarity) APG00030 (55.6% identity,65.9% similarity) APG00196 (52.5% identity, 64.2% similarity) AGP17990.1(51.3% identity, 61.3% similarity) APG00096 (51.2% identity, 62.5%similarity) US_8461415_B2-14_1 (50.4% identity, 61.0% similarity)AGP17989.1 (50.3% identity, 61.0% similarity) Cry70Ba1 (21.6% identity,34.1% similarity) APG00508 111 112 APG00355 (86.7% identity, 92.2%similarity) Cry 30, 35, 40, 45, 50, 55, 60, 65, 70, 40, 45, 50, 55, 60,65, 70, 75, 75, 80, 85, 90, 95, 96, 97, 98, 99 80, 85, 90, 95, 96, 97,98, 99 WP_017762616.1 (26.0% identity, 38.9% similarity) AGA40058.1(22.6% identity, 34.5% similarity) WP_017762581.1 (21.9% identity, 36.4%similarity) AGA40057.1 (21.7% identity, 34.5% similarity) APG00509 113285 APG00541 (98.4% identity, 99.0% similarity) Mtx 55, 60, 65, 70, 75,80, 85, 90, 95, 75, 80, 85, 90, 95, 96, 97, 98, 96, 97, 98, 99 99APG00244 (93.3% identity, 96.8% similarity) APG00896 (56.7% identity,74.0% similarity) WP_016078640.1 (53.4% identity, 72.5% similarity)WP_036654376.1 (47.3% identity, 60.8% similarity) WP_033699741.1 (46.2%identity, 63.5% similarity) ETK27180.1 (46.2% identity, 59.8%similarity) APG00510 114 286 APG00443 (72.5% identity, 72.9% similarity)Cry 40, 45, 50, 55, 60, 65, 70, 75, 80, 55, 60, 65, 70, 75, 80, 85, 90,85, 90, 95, 96, 97, 98, 99 95, 96, 97, 98, 99 WP_017762621.1 (39.7%identity, 52.7% similarity) WP_017762616.1 (25.8% identity, 41.4%similarity) AGA40058.1 (24.6% identity, 38.3% similarity) WP_017762619.1(22.8% identity, 36.3% similarity) APG00519 115 APG00381 (79.8%identity, 88.5% similarity) Cry6 80, 85, 90, 95, 96, 97, 98, 99 90, 95,96, 97, 98, 99 APG00397 (79.8% identity, 88.2% similarity) APG00720(78.4% identity, 87.5% similarity) WP_002144454.1 (78.0% identity, 86.4%similarity) WP_001072414.1 (77.7% identity, 85.7% similarity)WP_002187783.1 (75.6% identity, 85.4% similarity) WP_002169785.1 (62.3%identity, 76.5% similarity) APG00522 116 117 EFT94981.1 (39.0% identity,54.0% similarity) Mtx 40, 45, 50, 55, 60, 65, 70, 75, 80, 55, 60, 65,70, 75, 80, 85, 90, 85, 90, 95, 96, 97, 98, 99 95, 96, 97, 98, 99WP_010761370.1 (37.0% identity, 52.8% similarity) WP_010734336.1 (35.8%identity, 53.4% similarity) WP_016625081.1 (28.6% identity, 46.4%similarity) APG00535 118 APG00016 (84.4% identity, 90.4% similarity) Mtx30, 35, 40, 45, 50, 55, 60, 65, 70, 45, 50, 55, 60, 65, 70, 75, 80, 75,80, 85, 90, 95, 96, 97, 98, 99 85, 90, 95, 96, 97, 98, 99US20120278954A1_30 (28.9% identity, 43.5% similarity) Cry23Aa1 (28.4%identity, 43.1% similarity) APG00541 119 287 APG00509 (98.4% identity,99.0% similarity) Mtx 55, 60, 65, 70, 75, 80, 85, 90, 95, 75, 80, 85,90, 95, 96, 97, 98, 96, 97, 98, 99 99 APG00244 (94.6% identity, 97.4%similarity) APG00896 (57.1% identity, 74.0% similarity) WP_016078640.1(53.4% identity, 73.5% similarity) WP_036654376.1 (47.0% identity, 61.1%similarity) WP_033699741.1 (45.9% identity, 63.5% similarity) ETK27180.1(45.9% identity, 60.1% similarity) APG00542 120 121 WP_016099611.1(25.9% identity, 44.6% similarity) Mtx 30, 35, 40, 45, 50, 55, 60, 65,70, 45, 50, 55, 60, 65, 70, 75, 80, 75, 80, 85, 90, 95, 96, 97, 98, 9985, 90, 95, 96, 97, 98, 99 WP_016098287.1 (24.6% identity, 41.5%similarity) US_8318900_B2-78 (23.5% identity, 39.0% similarity)WP_033699741.1 (23.5% identity, 37.9% similarity) APG00545 122 288APG00276 (74.9% identity, 84.7% similarity) Mtx 35, 40, 45, 50, 55, 60,65, 70, 75, 50, 55, 60, 65, 70, 75, 80, 85, 80, 85, 90, 95, 96, 97, 98,99 90, 95, 96, 97, 98, 99 APG00671 (57.8% identity, 75.3% similarity)WP_000794513.1 (34.9% identity, 48.9% similarity) AGP18071.1 (34.4%identity, 48.9% similarity) WP_036654376.1 (32.8% identity, 45.9%similarity) ETK27180.1 (31.8% identity, 44.9% similarity) APG00546 123AGP17988.1 (46.7% identity, 66.4% similarity) Mtx 50, 55, 60, 65, 70,75, 80, 85, 90, 70, 75, 80, 85, 90, 95, 96, 97, 95, 96, 97, 98, 99 98,99 APG00553 124 APG00480 (69.1% identity, 84.3% similarity) Cry 25, 30,35, 40, 45, 50, 55, 60, 65, 40, 45, 50, 55, 60, 65, 70, 75, 70, 75, 80,85, 90, 95, 96, 97, 98, 80, 85, 90, 95, 96, 97, 98, 99 99 APG00733(59.4% identity, 75.3% similarity) APG00296 (50.3% identity, 65.1%similarity) US20130227743A1_200 (21.3% identity, 34.5% similarity)AGA40057.1 (21.1% identity, 33.4% similarity) WP_017762616.1 (20.1%identity, 35.8% similarity) WP_017762581.1 (20.1% identity, 33.4%similarity) APG00560 125 289 APG00269 (96.2% identity, 96.8% similarity)Mtx 55, 60, 65, 70, 75, 80, 85, 90, 95, 70, 75, 80, 85, 90, 95, 96, 97,96, 97, 98, 99 98, 99 APG00995 (85.0% identity, 89.9% similarity)APG00414 (83.9% identity, 89.4% similarity) APG00593 (83.9% identity,89.1% similarity) US20130227743A1_110 (53.6% identity, 68.5% similarity)AGP17985.1 (47.0% identity, 60.2% similarity) WP_002166885.1 (43.7%identity, 57.6% similarity) AGP17983.1 (43.6% identity, 59.2%similarity) APG00562 126 290 US20130227743A1_64 (25.1% identity, 42.4%Mtx 30, 35, 40, 45, 50, 55, 60, 65, 70, 45, 50, 55, 60, 65, 70, 75, 80,similarity) 75, 80, 85, 90, 95, 96, 97, 98, 99 85, 90, 95, 96, 97, 98,99 WP_000794514.1 (23.8% identity, 39.5% similarity) WP_000794513.1(23.1% identity, 42.6% similarity) AGP18071.1 (22.9% identity, 42.3%similarity) APG00563 127 291 APG00794 (85.5% identity, 91.4% similarity)Mtx 60, 65, 70, 75, 80, 85, 90, 95, 96, 75, 80, 85, 90, 95, 96, 97, 98,97, 98, 99 99 APG00569 (83.3% identity, 89.9% similarity) APG00938(82.7% identity, 91.1% similarity) APG00590 (59.8% identity, 74.9%similarity) APG00146 (57.9% identity, 70.9% similarity) WP_000794514.1(57.5% identity, 70.5% similarity) US20130227743A1_102 (56.7% identity,71.5% similarity) AGA40045.1 (55.4% identity, 70.4% similarity) APG00667(50.6% identity, 62.3% similarity) WP_036654376.1 (41.0% identity, 59.6%similarity) APG00564 128 292 AGA40058.1 (27.8% identity, 42.2%similarity) Cry 30, 35, 40, 45, 50, 55, 60, 65, 70, 45, 50, 55, 60, 65,70, 75, 80, 75, 80, 85, 90, 95, 96, 97, 98, 99 85, 90, 95, 96, 97, 98,99 AGA40057.1 (26.8% identity, 36.9% similarity) WP_017762619.1 (25.5%identity, 40.1% similarity) US_8461415_B2-45 (25.5% identity, 39.1%similarity) APG00569 129 293 APG00938 (88.4% identity, 94.0% similarity)Mtx 60, 65, 70, 75, 80, 85, 90, 95, 96, 75, 80, 85, 90, 95, 96, 97, 98,97, 98, 99 99 APG00563 (83.3% identity, 89.9% similarity) APG00794(77.4% identity, 87.2% similarity) APG00590 (58.4% identity, 75.8%similarity) APG00146 (58.0% identity, 71.9% similarity) WP_000794514.1(57.1% identity, 71.3% similarity) AGA40045.1 (56.7% identity, 70.3%similarity) US20130227743A1_102 (54.7% identity, 71.3% similarity)APG00667 (50.9% identity, 65.3% similarity) WP_036654376.1 (41.8%identity, 59.1% similarity) APG00570 130 WP_000794513.1 (29.4% identity,31.2% similarity) Mtx 30, 35, 40, 45, 50, 55, 60, 65, 70, 35, 40, 45,50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 75, 80, 85, 90,95, 96, 97, 98, 99 AGP18071.1 (29.1% identity, 30.9% similarity)ETK27180.1 (15.2% identity, 20.3% similarity) WP_016078640.1 (14.6%identity, 20.7% similarity) APG00579 131 294 WP_002166885.1 (26.4%identity, 38.7% similarity) Mtx 30, 35, 40, 45, 50, 55, 60, 65, 70, 45,50, 55, 60, 65, 70, 75, 80, 75, 80, 85, 90, 95, 96, 97, 98, 99 85, 90,95, 96, 97, 98, 99 US20130227743A1_110 (26.2% identity, 41.8%similarity) US_8318900_B2-75 (24.5% identity, 33.1% similarity)US_8829279_B2-11 (24.3% identity, 38.8% similarity) APG00580 132 295AGP17983.1 (36.0% identity, 50.1% similarity) Mtx 40, 45, 50, 55, 60,65, 70, 75, 80, 55, 60, 65, 70, 75, 80, 85, 90, 85, 90, 95, 96, 97, 98,99 95, 96, 97, 98, 99 AGP17985.1 (35.3% identity, 50.3% similarity)US20130227743A1_110 (32.9% identity, 48.9% similarity) US_8829279_B2-11(31.6% identity, 49.3% similarity) APG00581 133 134 AGP18059.1 (77.7%identity, 82.0% similarity) Cry62A 80, 85, 90, 95, 96, 97, 98, 99 85,90, 95, 96, 97, 98, 99 US_8461421_B2-45_1 (77.7% identity, 82.0%similarity) Cry62Aa1 (77.7% identity, 81.8% similarity) APG00586 135 296APG00420 (95.6% identity, 96.8% similarity) Mtx 40, 45, 50, 55, 60, 65,70, 75, 80, 55, 60, 65, 70, 75, 80, 85, 90, 85, 90, 95, 96, 97, 98, 9995, 96, 97, 98, 99 APG00323 (92.6% identity, 95.9% similarity)AGA40044.1 (35.8% identity, 54.5% similarity) WP_002203994.1 (16.8%identity, 19.2% similarity) APG00593 136 297 APG00995 (98.4% identity,98.9% similarity) Mtx 60, 65, 70, 75, 80, 85, 90, 95, 96, 75, 80, 85,90, 95, 96, 97, 98, 97, 98, 99 99 APG00414 (97.5% identity, 98.6%similarity) APG00560 (83.9% identity, 89.1% similarity) APG00269 (83.6%identity, 88.7% similarity) US20130227743A1_110 (56.0% identity, 71.6%similarity) AGP17985.1 (47.5% identity, 57.2% similarity) WP_002166885.1(45.0% identity, 58.9% similarity) AGP17983.1 (44.1% identity, 57.4%similarity) APG00594 137 298 APG00154 (98.0% identity, 98.6% similarity)Mtx 55, 60, 65, 70, 75, 80, 85, 90, 95, 70, 75, 80, 85, 90, 95, 96, 97,96, 97, 98, 99 98, 99 APG00486 (75.9% identity, 86.7% similarity)APG00421 (63.8% identity, 75.9% similarity) APG00483 (63.4% identity,76.2% similarity) US20130227743A1_110 (52.4% identity, 69.3% similarity)AGP17985.1 (46.2% identity, 60.4% similarity) AGP17983.1 (45.3%identity, 61.5% similarity) WP_002166885.1 (39.5% identity, 56.6%similarity) APG00595 138 299, 300 AEH76824.1 (85.7% identity, 91.5%similarity) Cry70 90, 95, 96, 97, 98, 99 95, 96, 97, 98, 99US_8318900_B2-119_1 (78.6% identity, 83.8% similarity) WP_016093954.1(66.8% identity, 79.3% similarity) Cry70Bb1 (65.6% identity, 78.6%similarity) APG00025 (65.1% identity, 76.7% similarity) APG00027 (53.9%identity, 69.4% similarity) APG00597 139 301 APG00268 (87.6% identity,91.3% similarity) Mtx 80, 85, 90, 95, 96, 97, 98, 99 85, 90, 95, 96, 97,98, 99 APG00764 (87.3% identity, 91.6% similarity) APG00393 (87.1%identity, 91.6% similarity) APG00481 (85.8% identity, 90.8% similarity)APG00635 (84.7% identity, 88.9% similarity) APG00020 (78.6% identity,84.7% similarity) WP_002166885.1 (76.3% identity, 83.1% similarity)APG00227 (55.9% identity, 66.8% similarity) US20130227743A1_110 (43.0%identity, 60.4% similarity) AGP17983.1 (36.5% identity, 54.6%similarity) AGP17985.1 (35.9% identity, 53.3% similarity) APG00598 140APG00236 (29.4% identity, 49.8% similarity) Mtx 25, 30, 35, 40, 45, 50,55, 60, 65, 25, 30, 35, 40, 45, 50, 55, 60, 70, 75, 80, 85, 90, 95, 96,97, 98, 65, 70, 75, 80, 85, 90, 95, 96, 99 97, 98, 99 APG00994 (19.5%identity, 34.9% similarity) APG00601 141 302 APG00021 (96.4% identity,98.9% similarity) Mtx 95, 96, 97, 98, 99 97, 98, 99 APG00091 (95.4%identity, 96.8% similarity) AGP18028.1 (90.7% identity, 96.1%similarity) APG00850 (75.0% identity, 86.4% similarity) Cry45Aa (32.2%identity, 47.3% similarity) US_8461421_B2-42_1 (32.2% identity, 47.1%similarity) APG00613 142 143 APG00004 (83.6% identity, 90.4% similarity)Cry 55, 60, 65, 70, 75, 80, 85, 90, 95, 70, 75, 80, 85, 90, 95, 96, 97,96, 97, 98, 99 98, 99 US_7605304_B2-8 (52.2% identity, 66.2% similarity)US_7605304_B2-7_1 (52.1% identity, 66.1% similarity) US_7105332_B2-14(51.7% identity, 65.6% similarity) Cry8Ja1 (34.0% identity, 41.7%similarity) APG00614 144 145 US20120278954A1_22 (90.0% identity, 93.1%Mtx 95, 96, 97, 98, 99 95, 96, 97, 98, 99 similarity) AF316145_1 (65.9%identity, 79.4% similarity) AAA22332.1 (32.8% identity, 47.6%similarity) WP_033694890.1 (29.9% identity, 41.0% similarity) APG00625146 303 AGP17992.1 (31.7% identity, 44.5% similarity) Cry 35, 40, 45,50, 55, 60, 65, 70, 75, 45, 50, 55, 60, 65, 70, 75, 80, 80, 85, 90, 95,96, 97, 98, 99 85, 90, 95, 96, 97, 98, 99 WP_017762581.1 (30.2%identity, 43.0% similarity) AGA40058.1 (29.7% identity, 43.8%similarity) Cry52Aa1 (13.3% identity, 24.3% similarity) APG00628 147 304APG00636 (76.2% identity, 86.4% similarity) Mtx 30, 35, 40, 45, 50, 55,60, 65, 70, 50, 55, 60, 65, 70, 75, 80, 85, 75, 80, 85, 90, 95, 96, 97,98, 99 90, 95, 96, 97, 98, 99 APG00834 (72.4% identity, 86.2%similarity) APG00634 (59.8% identity, 69.0% similarity) US_8829279_B2-11(29.6% identity, 45.8% similarity) AGP17984.1 (27.5% identity, 41.4%similarity) AGP18033.1 (26.2% identity, 39.0% similarity) AGS78124.1(25.0% identity, 40.6% similarity) APG00633 148 305 WP_016078640.1(51.0% identity, 69.3% similarity) Mtx 55, 60, 65, 70, 75, 80, 85, 90,95, 70, 75, 80, 85, 90, 95, 96, 97, 96, 97, 98, 99 98, 99 WP_033699741.1(45.9% identity, 60.6% similarity) WP_036654376.1 (44.1% identity, 54.8%similarity) WP_006284936.1 (43.8% identity, 59.4% similarity) APG00634149 306 APG00636 (63.3% identity, 71.1% similarity) Mtx 30, 35, 40, 45,50, 55, 60, 65, 70, 45, 50, 55, 60, 65, 70, 75, 80, 75, 80, 85, 90, 95,96, 97, 98, 99 85, 90, 95, 96, 97, 98, 99 APG00834 (61.2% identity,70.1% similarity) APG00628 (59.8% identity, 69.0% similarity)US_8829279_B2-11 (27.5% identity, 41.2% similarity) AGP17984.1 (25.7%identity, 37.9% similarity) AGP18033.1 (25.1% identity, 37.4%similarity) US20130227743A1_110 (23.8% identity, 36.5% similarity)APG00636 150 307 APG00834 (88.2% identity, 93.0% similarity) Mtx 35, 40,45, 50, 55, 60, 65, 70, 75, 50, 55, 60, 65, 70, 75, 80, 85, 80, 85, 90,95, 96, 97, 98, 99 90, 95, 96, 97, 98, 99 APG00628 (76.2% identity,86.4% similarity) APG00634 (63.3% identity, 71.1% similarity)US_8829279_B2-11 (31.2% identity, 47.2% similarity) AGP18033.1 (30.9%identity, 46.9% similarity) WP_000844425.1 (28.8% identity, 43.2%similarity) AGP17984.1 (28.8% identity, 42.4% similarity) APG00645 151APG00425 (71.8% identity, 80.2% similarity) Mtx 40, 45, 50, 55, 60, 65,70, 75, 80, 60, 65, 70, 75, 80, 85, 90, 95, 85, 90, 95, 96, 97, 98, 9996, 97, 98, 99 AGA40044.1 (38.3% identity, 58.5% similarity) APG00654152 153, 308 APG00164 (83.2% identity, 88.2% similarity) Cry 60, 65, 70,75, 80, 85, 90, 95, 96, 75, 80, 85, 90, 95, 96, 97, 98, 97, 98, 99 99CBL59393.1 (56.6% identity, 71.9% similarity) CBL59396.1 (56.6%identity, 71.9% similarity) D5H3I8_BACTG (56.5% identity, 71.8%similarity) APG00084 (52.5% identity, 68.2% similarity) Cry8Aa1 (36.0%identity, 42.7% similarity) APG00665 154 309 ETT84679.1 (36.8% identity,52.8% similarity) Cry 40, 45, 50, 55, 60, 65, 70, 75, 80, 55, 60, 65,70, 75, 80, 85, 90, 85, 90, 95, 96, 97, 98, 99 95, 96, 97, 98, 99WP_033728958.1 (36.4% identity, 52.3% similarity) US_2013_0227743_A1_196(35.6% identity, 53.6% similarity) Cry2Af1 (28.4% identity, 44.7%similarity) APG00667 155 310 APG00417 (62.3% identity, 73.0% similarity)Mtx 50, 55, 60, 65, 70, 75, 80, 85, 90, 70, 75, 80, 85, 90, 95, 96, 97,95, 96, 97, 98, 99 98, 99 APG00938 (52.2% identity, 64.2% similarity)APG00590 (51.4% identity, 67.9% similarity) APG00569 (50.9% identity,65.3% similarity) APG00794 (50.4% identity, 64.1% similarity)WP_000794514.1 (49.8% identity, 63.7% similarity) US20130227743A1_102(49.2% identity, 65.2% similarity) AGA40045.1 (46.6% identity, 61.9%similarity) ETK27180.1 (44.2% identity, 57.3% similarity) APG00669 156311 APG00731 (90.9% identity, 93.6% similarity) Bin 90, 95, 96, 97, 98,99 95, 96, 97, 98, 99 APG00231 (89.0% identity, 94.4% similarity)APG00035 (88.7% identity, 94.1% similarity) APG00377 (88.5% identity,94.1% similarity) APG00356 (88.2% identity, 93.6% similarity)WP_000143307.1 (87.9% identity, 92.8% similarity) APG00157 (86.6%identity, 93.3% similarity) WP_000143308.1 (85.5% identity, 92.8%similarity) US20130227743A1_6 (78.8% identity, 85.0% similarity)APG00257 (51.1% identity, 69.1% similarity) Cry35Ac1 (23.0% identity,42.6% similarity) APG00671 157 312 APG00545 (57.8% identity, 75.3%similarity) Mtx 35, 40, 45, 50, 55, 60, 65, 70, 75, 50, 55, 60, 65, 70,75, 80, 85, 80, 85, 90, 95, 96, 97, 98, 99 90, 95, 96, 97, 98, 99APG00276 (56.6% identity, 75.2% similarity) WP_036654376.1 (33.6%identity, 48.8% similarity) AGP18071.1 (32.9% identity, 48.2%similarity) ETK27180.1 (32.8% identity, 47.2% similarity) WP_000794513.1(32.4% identity, 48.0% similarity) APG00681 158 313 WP_030030026.1(62.6% identity, 74.7% similarity) Mtx 65, 70, 75, 80, 85, 90, 95, 96,97, 75, 80, 85, 90, 95, 96, 97, 98, 98, 99 99 APG00129 (62.6% identity,74.2% similarity) US20130227743A1_106 (20.6% identity, 36.2% similarity)APG00686 159 US_8829279_B2-37 (27.0% identity, 40.2% Mtx 30, 35, 40, 45,50, 55, 60, 65, 70, 45, 50, 55, 60, 65, 70, 75, 80, similarity) 75, 80,85, 90, 95, 96, 97, 98, 99 85, 90, 95, 96, 97, 98, 99 AGA40032.1 (25.8%identity, 40.7% similarity) AGA40044.1 (25.4% identity, 43.3%similarity) US_8829279_B2-27 (23.8% identity, 40.0% similarity) APG00696160 AGP18056.1 (25.3% identity, 41.9% similarity) Mtx 30, 35, 40, 45,50, 55, 60, 65, 70, 45, 50, 55, 60, 65, 70, 75, 80, 75, 80, 85, 90, 95,96, 97, 98, 99 85, 90, 95, 96, 97, 98, 99 US20120278954A1_26 (25.2%identity, 43.0% similarity) Cry45Aa (25.0% identity, 43.1% similarity)APG00704 161 314 APG00020 (81.9% identity, 87.9% similarity) Mtx 75, 80,85, 90, 95, 96, 97, 98, 99 80, 85, 90, 95, 96, 97, 98, 99 APG00764(79.6% identity, 84.3% similarity) APG00268 (78.3% identity, 83.5%similarity) APG00393 (78.2% identity, 84.0% similarity) APG00481 (77.5%identity, 82.7% similarity) APG00635 (76.4% identity, 83.0% similarity)WP_002166885.1 (73.8% identity, 79.0% similarity) APG00227 (53.9%identity, 63.7% similarity) US20130227743A1_110 (45.5% identity, 62.9%similarity) AGP17985.1 (37.0% identity, 54.9% similarity) AGP17983.1(33.2% identity, 51.2% similarity) APG00711 162 163 Cry5Ad1 (71.7%identity, 78.4% similarity) Cry5 75, 80, 85, 90, 95, 96, 97, 98, 99 80,85, 90, 95, 96, 97, 98, 99 APG00719 164 315, 316 WP_001036192.1 (46.9%identity, 64.0% similarity) Mtx 50, 55, 60, 65, 70, 75, 80, 85, 90, 65,70, 75, 80, 85, 90, 95, 96, 95, 96, 97, 98, 99 97, 98, 99 WP_003290257.1(45.0% identity, 63.5% similarity) WP_008180054.1 (43.5% identity, 62.4%similarity) WP_000790613.1 (43.3% identity, 62.2% similarity) APG00733165 APG00480 (71.4% identity, 80.8% similarity) Cry 25, 30, 35, 40, 45,50, 55, 60, 65, 40, 45, 50, 55, 60, 65, 70, 75, 70, 75, 80, 85, 90, 95,96, 97, 98, 80, 85, 90, 95, 96, 97, 98, 99 99 APG00553 (59.4% identity,75.3% similarity) AGA40057.1 (22.2% identity, 34.3% similarity)WP_017762616.1 (21.6% identity, 37.1% similarity) US20130227743A1_200(20.0% identity, 32.8% similarity) AGA40058.1 (18.8% identity, 30.7%similarity) APG00755 166 167 WP_002193629.1 (61.0% identity, 72.3%similarity) Mtx 65, 70, 75, 80, 85, 90, 95, 96, 97, 75, 80, 85, 90, 95,96, 97, 98, 98, 99 99 APG00402 (52.6% identity, 68.7% similarity)WP_011254649.1 (24.3% identity, 41.5% similarity) US20120278954A1_54(21.6% identity, 39.9% similarity) Q45728_BACTU (20.2% identity, 34.7%similarity) APG00764 168 317 APG00959 (90.2% identity, 94.1% similarity)Mtx 85, 90, 95, 96, 97, 98, 99 90, 95, 96, 97, 98, 99 APG00322 (89.6%identity, 93.4% similarity) APG00268 (88.3% identity, 92.3% similarity)APG00481 (88.3% identity, 92.3% similarity) APG00646 (87.0% identity,92.6% similarity) APG00635 (86.4% identity, 91.0% similarity) APG00020(83.8% identity, 88.3% similarity) WP_002166885.1 (81.9% identity, 85.9%similarity) APG00227 (56.0% identity, 66.7% similarity)US20130227743A1_110 (44.8% identity, 61.7% similarity) AGP17985.1 (35.8%identity, 51.4% similarity) AGP17983.1 (34.2% identity, 52.0%similarity) APG00780 169 WP_029295902.1 (62.0% identity, 76.0%similarity) Bin 65, 70, 75, 80, 85, 90, 95, 96, 97, 80, 85, 90, 95, 96,97, 98, 99 98, 99 WP_038445296.1 (57.8% identity, 74.7% similarity)WP_029977655.1 (46.1% identity, 63.0% similarity) WP_017478170.1 (43.0%identity, 58.4% similarity) APG00793 170 318, 319 US_8829279_B2-11(38.8% identity, 55.0% Mtx 40, 45, 50, 55, 60, 65, 70, 75, 80, 60, 65,70, 75, 80, 85, 90, 95, similarity) 85, 90, 95, 96, 97, 98, 99 96, 97,98, 99 WP_000844424.1 (32.1% identity, 49.6% similarity) WP_000844425.1(32.1% identity, 48.9% similarity) AGP17983.1 (31.3% identity, 48.7%similarity) APG00794 171 320 APG00563 (85.5% identity, 91.4% similarity)Mtx 60, 65, 70, 75, 80, 85, 90, 95, 96, 75, 80, 85, 90, 95, 96, 97, 98,97, 98, 99 99 APG00938 (78.9% identity, 88.1% similarity) APG00569(77.4% identity, 87.2% similarity) APG00590 (59.0% identity, 73.2%similarity) APG00146 (58.2% identity, 69.7% similarity) WP_000794514.1(57.6% identity, 69.7% similarity) US20130227743A1_102 (56.5% identity,71.0% similarity) AGA40045.1 (54.7% identity, 69.8% similarity) APG00667(50.4% identity, 64.1% similarity) WP_036654376.1 (42.0% identity, 59.2%similarity) APG00800 172 321 WP_012259841.1 (85.8% identity, 91.2%similarity) Cry 90, 95, 96, 97, 98, 99 95, 96, 97, 98, 99US20130227743A1_24 (84.6% identity, 88.3% similarity) WP_025988975.1(69.9% identity, 70.5% similarity) APG00088 (67.6% identity, 77.3%similarity) Cry8Ca1 (25.4% identity, 35.5% similarity) APG00820 173 322APG00560 (53.0% identity, 55.2% similarity) Mtx 40, 45, 50, 55, 60, 65,70, 75, 80, 50, 55, 60, 65, 70, 75, 80, 85, 85, 90, 95, 96, 97, 98, 9990, 95, 96, 97, 98, 99 APG00269 (52.2% identity, 54.4% similarity)APG00593 (50.0% identity, 56.7% similarity) US20130227743A1_110 (35.7%identity, 48.8% similarity) AGP17985.1 (33.7% identity, 43.5%similarity) AGP17983.1 (32.6% identity, 44.7% similarity) EJQ15321.1(27.8% identity, 45.3% similarity) APG00825 174 323 APG00272 (79.5%identity, 90.4% similarity) Mtx 45, 50, 55, 60, 65, 70, 75, 80, 85, 60,65, 70, 75, 80, 85, 90, 95, 90, 95, 96, 97, 98, 99 96, 97, 98, 99APG00019 (66.3% identity, 75.7% similarity) WP_034679607.1 (41.2%identity, 59.4% similarity) WP_001039209.1 (35.3% identity, 53.5%similarity) US_8829279_B2-2 (34.1% identity, 52.6% similarity)US_8829279_B2-61 (33.6% identity, 52.0% similarity) APG00833 175 324APG00143 (80.4% identity, 87.6% similarity) Mtx 35, 40, 45, 50, 55, 60,65, 70, 75, 55, 60, 65, 70, 75, 80, 85, 90, 80, 85, 90, 95, 96, 97, 98,99 95, 96, 97, 98, 99 APG00441 (79.4% identity, 86.9% similarity)APG00444 (78.8% identity, 86.3% similarity) WP_006918908.1 (32.0%identity, 51.1% similarity) APG00834 176 325 APG00636 (88.2% identity,93.0% similarity) Mtx 35, 40, 45, 50, 55, 60, 65, 70, 75, 50, 55, 60,65, 70, 75, 80, 85, 80, 85, 90, 95, 96, 97, 98, 99 90, 95, 96, 97, 98,99 APG00628 (72.4% identity, 86.2% similarity) APG00634 (61.2% identity,70.1% similarity) US_8829279_B2-11 (31.2% identity, 46.2% similarity)AGP17984.1 (30.4% identity, 44.2% similarity) AGP18033.1 (30.2%identity, 43.8% similarity) WP_000844425.1 (27.6% identity, 39.8%similarity) APG00846 177 AGA40030.1 (62.1% identity, 77.4% similarity)Mtx 65, 70, 75, 80, 85, 90, 95, 96, 97, 80, 85, 90, 95, 96, 97, 98, 9998, 99 US_8829279_B2-37 (60.3% identity, 74.3% similarity)US_8829279_B2-27 (53.8% identity, 67.8% similarity) APG00224 (52.8%identity, 70.9% similarity) CAA67205.1 (43.7% identity, 60.9%similarity) APG00850 178 AGP18028.1 (77.1% identity, 88.2% similarity)Mtx 80, 85, 90, 95, 96, 97, 98, 99 90, 95, 96, 97, 98, 99 APG00601(75.0% identity, 86.4% similarity) APG00021 (74.3% identity, 87.5%similarity) APG00091 (73.9% identity, 86.1% similarity) Cry45Aa (31.6%identity, 48.6% similarity) APG00854 179 326 APG00911 (88.3% identity,93.6% similarity) Mtx 60, 65, 70, 75, 80, 85, 90, 95, 96, 75, 80, 85,90, 95, 96, 97, 98, 97, 98, 99 99 US_8829279_B2-11 (56.8% identity,72.6% similarity) APG00293 (50.9% identity, 64.0% similarity) APG00051(50.5% identity, 64.5% similarity) WP_000844424.1 (38.1% identity, 51.1%similarity) WP_000844425.1 (37.8% identity, 50.8% similarity)WP_033690552.1 (33.8% identity, 45.3% similarity) APG00860 180AGA40044.1 (41.2% identity, 55.2% similarity) Mtx 45, 50, 55, 60, 65,70, 75, 80, 85, 60, 65, 70, 75, 80, 85, 90, 95, 90, 95, 96, 97, 98, 9996, 97, 98, 99 APG00861 181 182 AGU13849.1 (63.6% identity, 72.4%similarity) Cry8 65, 70, 75, 80, 85, 90, 95, 96, 97, 75, 80, 85, 90, 95,96, 97, 98, 98, 99 99 Cry8Aa1 (60.3% identity, 71.1% similarity)APG00866 183 US20120278954A1_26 (48.7% identity, 68.4% Mtx 50, 55, 60,65, 70, 75, 80, 85, 90, 70, 75, 80, 85, 90, 95, 96, 97, similarity) 95,96, 97, 98, 99 98, 99 Cry33Aa1 (43.5% identity, 61.6% similarity)APG00891 184 327, 328 APG00426 (93.9% identity, 94.6% similarity) Mtx55, 60, 65, 70, 75, 80, 85, 90, 95, 65, 70, 75, 80, 85, 90, 95, 96, 96,97, 98, 99 97, 98, 99 APG00227 (93.5% identity, 95.0% similarity)APG00517 (78.5% identity, 80.3% similarity) APG00158 (61.8% identity,69.3% similarity) APG00481 (57.5% identity, 68.6% similarity) APG00597(56.6% identity, 68.4% similarity) APG00635 (54.7% identity, 66.0%similarity) APG00020 (53.9% identity, 65.7% similarity) WP_002166885.1(51.1% identity, 61.6% similarity) US20130227743A1_110 (40.9% identity,55.4% similarity) AGP17985.1 (35.1% identity, 50.5% similarity)US_8829279_B2-11 (31.0% identity, 45.0% similarity) APG00896 185 329APG00244 (58.0% identity, 73.7% similarity) Mtx 50, 55, 60, 65, 70, 75,80, 85, 90, 70, 75, 80, 85, 90, 95, 96, 97, 95, 96, 97, 98, 99 98, 99APG00541 (57.1% identity, 74.0% similarity) APG00509 (56.7% identity,74.0% similarity) WP_016078640.1 (49.5% identity, 67.0% similarity)WP_036654376.1 (47.8% identity, 66.9% similarity) ETK27180.1 (47.5%identity, 66.4% similarity) WP_006284936.1 (45.3% identity, 65.6%similarity) APG00897 186 187, 330, 331 Cry42Aa1 (46.4% identity, 56.3%similarity) Cry42 50, 55, 60, 65, 70, 75, 80, 85, 90, 60, 65, 70, 75,80, 85, 90, 95, 95, 96, 97, 98, 99 96, 97, 98, 99 APG00899 188 APG00098(98.6% identity, 98.8% similarity) Mtx 85, 90, 95, 96, 97, 98, 99 90,95, 96, 97, 98, 99 AGA40063.1 (80.4% identity, 85.9% similarity)AGA40061.1 (35.5% identity, 50.8% similarity) US_8318900_B2-191_1 (33.8%identity, 47.5% similarity) Cry60Ba2 (30.7% identity, 44.7% similarity)APG00911 189 332 APG00854 (88.3% identity, 93.6% similarity) Mtx 60, 65,70, 75, 80, 85, 90, 95, 96, 75, 80, 85, 90, 95, 96, 97, 98, 97, 98, 9999 US_8829279_B2-11 (58.4% identity, 73.6% similarity) APG00293 (51.1%identity, 65.5% similarity) WP_000844424.1 (36.9% identity, 50.7%similarity) WP_000844425.1 (36.6% identity, 50.4% similarity) AGP17983.1(33.4% identity, 48.7% similarity) APG00938 190 333 APG00569 (88.4%identity, 94.0% similarity) Mtx 60, 65, 70, 75, 80, 85, 90, 95, 96, 75,80, 85, 90, 95, 96, 97, 98, 97, 98, 99 99 APG00563 (82.7% identity,91.1% similarity) APG00794 (78.9% identity, 88.1% similarity) APG00590(59.4% identity, 75.1% similarity) APG00146 (58.3% identity, 71.4%similarity) AGA40045.1 (57.4% identity, 70.7% similarity) WP_000794514.1(57.4% identity, 70.4% similarity) US20130227743A1_102 (55.5% identity,72.1% similarity) APG00667 (52.2% identity, 64.2% similarity)WP_036654376.1 (43.8% identity, 61.2% similarity) APG00940 191 334AGP17988.1 (14.6% identity, 25.4% similarity) Mtx 15, 20, 25, 30, 35,40, 45, 50, 55, 30, 35, 40, 45, 50, 55, 60, 65, 60, 65, 70, 75, 80, 85,90, 95, 96, 70, 75, 80, 85, 90, 95, 96, 97, 97, 98, 99 98, 99 APG00950192 193 ACI01644.1 (24.3% identity, 39.2% similarity) Cry 25, 30, 35,40, 45, 50, 55, 60, 65, 40, 45, 50, 55, 60, 65, 70, 75, 70, 75, 80, 85,90, 95, 96, 97, 98, 80, 85, 90, 95, 96, 97, 98, 99 99 AGP17982.1 (22.2%identity, 37.1% similarity) Cry21Aa2 (19.1% identity, 32.5% similarity)APG00959 194 335, 336 APG00764 (90.2% identity, 94.1% similarity) Mtx80, 85, 90, 95, 96, 97, 98, 99 85, 90, 95, 96, 97, 98, 99 APG00322(89.3% identity, 91.7% similarity) APG00393 (89.0% identity, 92.5%similarity) APG00481 (88.8% identity, 91.5% similarity) APG00635 (85.6%identity, 89.6% similarity) APG00020 (79.5% identity, 85.4% similarity)WP_002166885.1 (77.9% identity, 84.0% similarity) APG00227 (56.1%identity, 66.7% similarity) US20130227743A1_110 (44.6% identity, 61.7%similarity) AGP17985.1 (36.0% identity, 52.2% similarity) AGP17983.1(35.1% identity, 53.6% similarity) APG00966 195 337, 338 APG00283 (57.9%identity, 72.6% similarity) Mtx 35, 40, 45, 50, 55, 60, 65, 70, 75, 55,60, 65, 70, 75, 80, 85, 90, 80, 85, 90, 95, 96, 97, 98, 99 95, 96, 97,98, 99 APG00479 (54.0% identity, 68.9% similarity) APG00411 (52.2%identity, 64.3% similarity) AGS78124.1 (34.9% identity, 53.8%similarity) AGP17984.1 (33.9% identity, 51.0% similarity)US_8829279_B2-11 (31.8% identity, 50.9% similarity) AGP18033.1 (31.6%identity, 49.9% similarity) APG00971 196 197, 339, 340 Cry7Ea3 (42.5%identity, 59.8% similarity) Cry 45, 50, 55, 60, 65, 70, 75, 80, 85, 60,65, 70, 75, 80, 85, 90, 95, 90, 95, 96, 97, 98, 99 96, 97, 98, 99APG00994 198 199 WP_033694378.1 (46.2% identity, 56.3% similarity) Mtx50, 55, 60, 65, 70, 75, 80, 85, 90, 60, 65, 70, 75, 80, 85, 90, 95, 95,96, 97, 98, 99 96, 97, 98, 99 US20060191034A1_33 (24.3% identity, 41.6%similarity) WP_019419944.1 (24.2% identity, 39.7% similarity)US20060191034A1_10 (23.8% identity, 42.1% similarity) APG00995 200 341APG00593 (98.4% identity, 98.9% similarity) Mtx 60, 65, 70, 75, 80, 85,90, 95, 96, 75, 80, 85, 90, 95, 96, 97, 98, 97, 98, 99 99 APG00414(98.1% identity, 98.6% similarity) APG00560 (85.0% identity, 89.9%similarity) APG00269 (84.7% identity, 89.5% similarity)US20130227743A1_110 (55.5% identity, 71.3% similarity) AGP17985.1 (47.0%identity, 56.7% similarity) WP_002166885.1 (45.0% identity, 58.9%similarity) AGP17983.1 (42.8% identity, 56.9% similarity) APG00646 201202, 203 APG00268 (93.4% identity, 96.5% similarity) Mtx 85, 90, 95, 96,97, 98, 99 90, 95, 96, 97, 98, 99 APG00481 (92.3% identity, 95.8%similarity) APG00322 (89.1% identity, 92.8% similarity) APG00393 (87.5%identity, 92.8% similarity) APG00635 (87.2% identity, 91.2% similarity)APG00764 (87.0% identity, 92.6% similarity) APG00020 (83.2% identity,87.8% similarity) WP_002166885.1 (80.1% identity, 85.4% similarity)WP_063226258.1 (48.4% identity, 62.2% similarity) US_2013_0227743_A1-110(44.0% identity, 60.4% similarity)

i. Classes of Pesticidal Proteins

The pesticidal proteins provided herein and the nucleotide sequencesencoding them are useful in methods for impacting pests. That is, thecompositions and methods of the invention find use in agriculture forcontrolling or killing pests, including pests of many crop plants. Thepesticidal proteins provided herein are toxin proteins from bacteria andexhibit activity against certain pests. The pesticidal proteins are fromseveral classes of toxins including Cry, Cyt, BIN, and Mtx toxins. See,for example, Table 1 for the specific protein classifications of thevarious SEQ ID NOS provided herein. In addition, reference is madethroughout this disclosure to Pfam database entries. The Pfam databaseis a database of protein families, each represented by multiple sequencealignments and a profile hidden Markov model. Finn et al. (2014) Nucl.Acid Res. Database Issue 42:D222-D230.

Bacillus thuringiensis (Bt) is a gram-positive bacterium that producesinsecticidal proteins as crystal inclusions during its sporulation phaseof growth. The is proteinaceous inclusions of Bacillus thuringiensis(Bt) are called crystal proteins or δ-endotoxins (or Cry proteins),which are toxic to members of the class Insecta and other invertebrates.Similarly, Cyt proteins are parasporal inclusion proteins from Bt thatexhibits hemolytic (Cytolytic) activity or has obvious sequencesimilarity to a known Cyt protein. These toxins are highly specific totheir target organism, and are innocuous to humans, vertebrates, andplants.

The structure of the Cry toxins reveals five conserved amino acidblocks, concentrated mainly in the center of the domain or at thejunction between the domains. The Cry toxin consists of three domains,each with a specific function. Domain I is a seven α-helix bundle inwhich a central helix is completely surrounded by six outer helices.This domain is implicated in channel formation in the membrane. DomainII appears as a triangular column of three anti-parallel β-sheets, whichare similar to antigen-binding regions of immunoglobulins. Domain IIIcontains anti-parallel β-strands in a β sandwich form. The N-terminalpart of the toxin protein is responsible for its toxicity andspecificity and contains five conserved regions. The C-terminal part isusually highly conserved and probably responsible for crystal formation.See, for example, U.S. Pat. No. 8,878,007.

Strains of B. thuringiensis show a wide range of specificity againstdifferent insect orders (Lepidoptera, Diptera, Coleoptera, Hymenoptera,Homoptera, Phthiraptera or Mallophaga, and Acari) and otherinvertebrates (Nemathelminthes, Platyhelminthes, and Sarocomastebrates).The Cry proteins have been classified into groups based on toxicity tovarious insect and invertebrate groups. Generally, Cry I demonstratestoxicity to lepidopterans, Cry II to lepidopterans and dipterans, CryIIIto coleopterans, Cry IV to dipterans, and Cry V and Cry VI to nematodes.New Cry proteins can be identified and assigned to a Cry group based onamino acid identity. See, for example, Bravo, A. (1997) J. of Bacteriol.179:2793-2801; Bravo et al. (2013) Microb. Biotechnol. 6:17-26, hereinincorporated by reference.

Over 750 different cry gene sequences have been classified into 73groups (Cry1-Cry73), with new members of this gene family continuing tobe discovered (Crickmore et al. (2014) www.btnomenclature.info/). Thecry gene family consists of several phylogentically non-related proteinfamilies that may have different modes of action: the family ofthree-domain Cry toxins, the family of mosquitocidal Cry toxins, thefamily of the binary-like toxins, and the Cyt family of toxins (Bravo etal., 2005). Some Bt strains produce additional insecticidal toxins, theVIP toxins. See, also, Cohen et al. (2011) J. Mol. Biol. 413:4-814;Crickmore et al. (2014) Bacillus thuringiensis toxin nomenclature, foundon the world wide web at lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/;Crickmore et al. (1988) Microbiol. Mol. Biol. Rev. 62: 807-813; Gill etal. (1992) Ann. Rev. Entomol. 37: 807-636; Goldbert et al. (1997) Appl.Environ. Microbiol. 63:2716-2712; Knowles et al. (1992) Proc. R. Soc.Ser. B. 248: 1-7; Koni et al. (1994) Microbiology 140: 1869-1880; Lailaket al. (2013) Biochem. Biophys. Res. Commun. 435: 216-221; Lopez-Diaz etal. (2013) Environ. Microbiol. 15: 3030-3039; Perez et al. (2007) Cell.Microbiol. 9: 2931-2937; Promdonkoy et al. (2003) Biochem. J. 374:255-259; Rigden (2009) FEBS Lett. 583: 1555-1560; Schnepf et al. (1998)Microbiol. Mol. Biol. Rev. 62: 775-806; Soberon et al. (2013) Peptides41: 87-93; Thiery et al. (1998) J. Am. Mosq. Control Assoc. 14: 472-476;Thomas et al. (1983) FEBS Lett. 154: 362-368; Wirth et al. (1997) Proc.Natl. Acad. Sci. U.S.A. 94: 10536-10540; Wirth et al (2005) Appl.Environ. Microbiol. 71: 185-189; and, Zhang et al. (2006) Biosci.Biotechnol. Biochem. 70: 2199-2204; each of which is herein incorporatedby reference in their entirety.

Cyt designates a parasporal crystal inclusion protein from Bacillusthuringiensis with cytolytic activity, or a protein with sequencesimilarity to a known Cyt protein. (Crickmore et al. (1998) Microbiol.Mol. Biol. Rev. 62: 807-813). The gene is denoted by cyt. These proteinsare different in structure and activity from Cry proteins (Gill et al.(1992) Annu. Rev. Entomol. 37: 615-636). The Cyt toxins were firstdiscovered in B. thuringiensis subspecies israelensis (Goldberg et al.(1977) Mosq. News. 37: 355-358). There are 3 Cyt toxin familiesincluding 11 holotype toxins in the current nomenclature (Crickmore etal. (2014) Bacillus thuringiensis toxin nomenclature found on the worldwide web at lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/). The majorityof the B. thuringiensis isolates with cyt genes show activity againstdipteran insects (particularly mosquitoes and black flies), but thereare also cyt genes that have been described in B. thuringiensis strainstargeting lepidopteran or coleopteran insects (Guerchicoff et al. (1997)Appl. Environ. Microbiol. 63: 2716-2721).

The structure of Cyt2A, solved by X-ray crystallography, shows a singledomain where two outer layers of α-helix wrap around a mixed β-sheet.Further available crystal structures of Cyt toxins support a conservedα-β structural model with two α-helix hairpins flanking a β-sheet corecontaining seven to eight β-strands. (Cohen et al. (2011) J. Mol. Biol.413: 80 4-814) Mutagenic studies identified β-sheet residues as criticalfor toxicity, while mutations in the helical domains did not affecttoxicity (Adang et al.; Diversity of Bacillus thuringiensis CrystalToxins and Mechanism of Action. In: T. S. Dhadialla and S. S. Gill, eds,Advances in Insect Physiology, Vol. 47, Oxford: Academic Press, 2014,pp. 39-87.) The representative domain of the Cyt toxin is a δ-endotoxin,Bac_thur_toxin (Pfam PF01338).

There are multiple proposed models for the mode of action of Cyt toxins,and it is still an area of active investigation. Some Cyt proteins(Cyt1A) have been shown to require the presence of accessory proteinsfor crystallization. Cyt1A and Cyt2A protoxins are processed bydigestive proteases at the same sites in the N- and C-termini to astable toxin core. Cyt toxins then interact with non-saturated membranelipids, such as phosphatidylcholine, phosphatidylethanolamine, andsphingomyelin. For Cyt toxins, pore-formation and detergent-likemembrane disruption have been proposed as non-exclusive mechanisms; andit is generally accepted that both may occur depending on toxinconcentration, with lower concentrations favoring oligomeric pores andhigher concentrations leading to membrane breaks. (Butko (2003) Appl.Environ. Microbiol. 69: 2415-2422) In the pore-formation model, the Cyttoxin binds to the cell membrane, inducing the formation ofcation-selective channels in the membrane vesicles leading tocolloid-osmotic lysis of the cell. (Knowles et al. (1989) FEBS Lett.244: 259-262; Knowles et al. (1992) Proc. R. Soc. Ser. B. 248: 1-7 andPromdonkoy et al. (2003) Biochem. J. 374: 255-259). In the detergentmodel, there is a nonspecific aggregation of the toxin on the surface ofthe lipid bilayer leading to membrane disassembly and cell death. (Butko(2003), supra; Manceva et al. (2005) Biochem. 44: 589-597).

Multiple studies have shown synergistic activity between Cyt toxins andother B. thuringiensis toxins, particularly the Cry, Bin, and Mtxtoxins. This synergism has even been shown to overcome an insect'sresistance to the other toxin. (Wirth 1997, Wirth 2005, Thiery 1998,Zhang 2006) The Cyt synergistic effect for Cry toxins is proposed toinvolve Cyt1A binding to domain II of Cry toxins in solution or on themembrane plane to promote formation of a Cry toxin pre-pore oligomer.Formation of this oligomer is independent of the Cyt oligomerization,binding, or insertion. (Lailak 2013, Perez 2007, Lopez-Diaz 2013)

A number of pesticidal proteins unrelated to the Cry proteins areproduced by some strains of B. thuringiensis and B. cereus duringvegetative growth (Estruch et al. (1996) Proc Natl Acad Sci USA93:5389-5394; Warren et al. (1994) WO 94/21795). These vegetativeinsecticidal proteins, or Vips, do not form parasporal crystal proteinsand are apparently secreted from the cell. The Vips are presentlyexcluded from the Cry protein nomenclature because they are notcrystal-forming proteins. The term VIP is a misnomer in the sense thatsome B. thuringiensis Cry proteins are also produced during vegetativegrowth as well as during the stationary and sporulation phases, mostnotably Cry3Aa. The location of the Vip genes in the B. thuringiensisgenome has been reported to reside on large plasmids that also encodecry genes (Mesrati et al. (2005) FEMS Microbiol. Lett. 244(2):353-8). Aweb-site for the nomenclature of Bt toxins can be found on the worldwide web at lifesci.sussex.ac.uk with the path“/home/Neil_Crickmore/Bt/” and at: “btnomenclature.info/”. See also,Schnepf et al. (1998) Microbiol. Mol. Biol. Rev. 62(3):775-806. Suchreferences are herein incorporated by reference.

To date four categories of Vips have been identified. Some Vip genesform binary two-component protein complexes; an “A” component is usuallythe “active” portion, and a “B” component is usually the “binding”portion. (Pfam pfam.xfam.org/family/PF03495). The Vip1 and Vip4 proteinsgenerally contain binary toxin B protein domains. Vip2 proteinsgenerally contain binary toxin A protein domains.

The Vip1 and Vip2 proteins are the two components of a binary toxin thatexhibits toxicity to coleopterans. Vip1Aa1 and Vip2Aa1 are very activeagainst corn rootworms, particularly Diabrotica virgifera and Diabroticalongicornis (Han et al. (1999) Nat. Struct. Biol. 6:932-936; Warren G W(1997) “Vegetative insecticidal proteins: novel proteins for control ofcorn pests” In: Carozzi N B, Koziel M (eds) Advances in insect control,the role of transgenic plants; Taylor & Francis Ltd, London, pp 109-21).The membrane-binding 95 kDa Vip1 multimer provides a pathway for the 52kDa Vip2 ADP-ribosylase to enter the cytoplasm of target western cornrootworm cells (Warren (1997) supra). The NAD-dependentADP-ribosyltransferase Vip2 likely modifies monomeric actin at Arg177 toblock polymerization, leading to loss of the actin cytoskeleton andeventual cell death due to the rapid subunit ex-change within actinfilaments in vivo (Carlier M. F. (1990) Adv. Biophys. 26:51-73).

Like Cry toxins, activated Vip3A toxins are pore-forming proteinscapable of making stable ion channels in the membrane (Lee et al. (2003)Appl. Environ. Microbiol. 69:4648-4657). Vip3 proteins are activeagainst several major lepidopteran pests (Rang et al. (2005) Appl.Environ. Microbiol. 71(10):6276-6281; Bhalla et al. (2005) FEMSMicrobiol. Lett. 243:467-472; Estruch et al. (1998) WO 9844137; Estruchet al. (1996) Proc Natl Acad Sci USA 93:5389-5394; Selvapandiyan et al.(2001) Appl. Environ Microbiol. 67:5855-5858; Yu et al. (1997) Appl.Environ Microbiol. 63:532-536). Vip3A is active against Agrotis ipsilon,Spodoptera frugiperda, Spodoptera exigua, Heliothis virescens, andHelicoverpa zea (Warren et al. (1996) WO 96/10083; Estruch et al. (1996)Proc Natl Acad Sci USA 93:5389-5394). Like Cry toxins, Vip3A proteinsmust be activated by proteases prior to recognition at the surface ofthe midgut epithelium of specific membrane proteins different from thoserecognized by Cry toxins.

The MTX family of toxin proteins is characterized by the presence of aconserved domain, ETX_MTX2 (pfam 03318). Members of this family sharesequence homology with the mosquitocidal toxins Mtx2 and Mtx3 fromBacillus sphaericus, as well as with the epsilon toxin ETX fromClostridium perfringens (Cole et al. (2004) Nat. Struct. Mol. Biol. 11:797-8; Thanabalu et al. (1996) Gene 170:85-9). The MTX-like proteins arestructurally distinct from the three-domain Cry toxins, as they have anelongated and predominately β-sheet-based structure. However, similar tothe three-domain toxins, the MTX-like proteins are thought to form poresin the membranes of target cells (Adang et al. (2014) supra). Unlike thethree-domain Cry proteins, the MTX-like proteins are much smaller inlength, ranging from 267 amino acids (Cry23) to 340 amino acids(Cry15A).

To date, only 15 proteins belonging to the family of MTX-like toxinshave been assigned Cry names, making this a relatively small classcompared to the three-domain Cry family (Crickmore et al. (2014) supra;Adang et al. (2014) supra). The members of the MTX-like toxin familyinclude Cry15, Cry23, Cry33, Cry38, Cry45, Cry46, Cry51, Cry60A, Cry60B,and Cry64. This family exhibits a range of insecticidal activity,including activity against insect pests of the Lepidopteran andColeopteran orders. Some members of this family may form binarypartnerships with other proteins, which may or may not be required forinsecticidal activity.

Cry15 is a 34 kDA protein that was identified in Bacillus thuringiensisserovar thompsoni HD542; it occurs naturally in a crystal together withan unrelated protein of approximately 40 kDa. The gene encoding Cry15and its partner protein are arranged together in an operon. Cry15 alonehas been shown to have activity against lepidopteran insect pestsincluding Manduca sexta, Cydia pomonella, and Pieris rapae, with thepresence of the 40 kDA protein having been shown to increase activity ofCry15 only against C. pomonella (Brown K. and Whiteley H. (1992) J.Bacteriol. 174:549-557; Naimov et al. (2008) Appl. Environ. Microbiol.74:7145-7151). Further studies are needed to elucidate the function ofthe partner protein of Cry15. Similarly, Cry23 is a 29 kDA protein thathas been shown to have activity against the coleopteran pests Triboliumcastaneum and Popillia japonica together with its partner protein Cry37(Donovan et al. (2000); U.S. Pat. No. 6,063,756).

New members of the MTX-like family are continuing to be identified. AnETX_MTX toxin gene was recently identified in the genome of Bacillusthuringiensis serovar tolworthi strain Na205-3. This strain was found tobe toxic against the lepidpoteran pest Helicoverpa armigera, and it alsocontained homologs of Cry 1, Cry 11, Vip1, Vip2, and Vip3 (Palma et al.(2014) Genome Announc. 2(2): e00187-14. Published online Mar. 13, 2014at doi: 10.1128/genomeA.00187-14; PMCID: PMC3953196). Because theMTX-like proteins have a unique domain structure relative to thethree-domain Cry proteins, they are believed to possess a unique mode ofaction, thereby making them a valuable tool in insect control and thefight against insect resistance.

Bacterial cells produce large numbers of toxins with diverse specificityagainst host and non-host organisms. Large families of binary toxinshave been identified in numerous bacterial families, including toxinsthat have activity against insect pests. (Poopathi and Abidha (2010) J.Physiol. Path. 1(3): 22-38). Lysinibacillus sphaericus (Ls), formerlyBacillus sphaericus, (Ahmed et al. (2007) Int. J Syst. Evol. Microbiol.57:1117-1125) is well-known as an insect biocontrol strain. Ls producesseveral insecticidal proteins, including the highly potent binarycomplex BinA/BinB. This binary complex forms a parasporal crystal in Lscells and has strong and specific activity against dipteran insects,specifically mosquitos. In some areas, insect resistance to existing Lsmosquitocidal strains has been reported. The discovery of new binarytoxins with different target specificity or the ability to overcomeinsect resistance is of significant interest.

The Ls binary insecticidal protein complex contains two majorpolypeptides, a 42 kDa polypeptide and a 51 kDa polypepdide, designatedBinA and BinB, respectively (Ahmed et al. (2007) supra). The twopolypeptides act synergistically to confer toxicity to their targets.Mode of action involves binding of the proteins to receptors in thelarval midgut. In some cases, the proteins are modified by proteasedigestion in the larval gut to produce activated forms. The BinBcomponent is thought to be involved in binding, while the BinA componentconfers toxicity (Nielsen-LeRoux et al. (2001) Appl. Environ. Microbiol.67(11):5049-5054). When cloned and expressed separately, the BinAcomponent is toxic to mosquito larvae, while the BinB component is not.However, co-administration of the proteins markedly increases toxicity(Nielsen-LeRoux et al. (2001) supra).

A small number of Bin protein homologs have been described frombacterial sources. Priest et al. (1997) Appl. Environ. Microbiol.63(4):1195-1198 describe a hybridization effort to identify new Lsstrains, although most of the genes they identified encoded proteinsidentical to the known BinA/BinB proteins. The BinA protein contains adefined conserved domain known as the Toxin 10 superfamily domain. Thistoxin domain was originally defined by its presence in BinA and BinB.The two proteins both have the domain, although the sequence similaritybetween BinA and BinB is limited in this region (<40%). The Cry49Aaprotein, which also has insecticidal activity, also has this domain(described below).

The Cry48Aa/Cry49Aa binary toxin of Ls has the ability to kill Culexquinquefasciatus mosquito larvae. These proteins are in a proteinstructural class that has some similarity to the Cry protein complex ofBacillus thuringiensis (Bt), a well-known insecticidal protein family.The Cry34/Cry35 binary toxin of Bt is also known to kill insects,including Western corn rootworm, a significant pest of corn. Cry34, ofwhich several variants have been identified, is a small (14 kDa)polypeptide, while Cry35 (also encoded by several variants) is a 44 kDapolypeptide. These proteins have some sequence homology with theBinA/BinB protein group and are thought to be evolutionarily related(Ellis et al. (2002) Appl. Environ. Microbiol. 68(3):1137-1145).

Provided herein are pesticidal proteins from these classes of toxins.The pesticidal proteins are classified by their structure, homology toknown toxins and/or their pesticidal specificity.

ii. Variants and Fragments of Pesticidal Proteins and PolynucleotidesEncoding the Same

Pesticidal proteins or polypeptides of the invention include those setforth in SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51,52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104,105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118,119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132,133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146,147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160,161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174,175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188,189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202,203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216,217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230,231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244,245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258,259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272,273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286,287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300,301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314,315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328,329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, and/or341and fragments and variants thereof. By “pesticidal toxin” or“pesticidal protein” or “pesticidal polypeptide” is intended a toxin orprotein or polypeptide that has activity against one or more pests,including, insects, fungi, nematodes, and the like such that the pest iskilled or controlled.

An “isolated” or “purified” polypeptide or protein, or biologicallyactive portion thereof, is substantially or essentially free fromcomponents that normally accompany or interact with the polypeptide orprotein as found in its naturally occurring environment. Thus, anisolated or purified polypeptide or protein is substantially free ofother cellular material, or culture medium when produced by recombinanttechniques, or substantially free of chemical precursors or otherchemicals when chemically synthesized. A protein that is substantiallyfree of cellular material includes preparations of protein having lessthan about 30%, 20%, 10%, 5%, or 1% (by dry weight) of contaminatingprotein. When the protein of the invention or biologically activeportion thereof is recombinantly produced, optimally culture mediumrepresents less than about 30%, 20%, 10%, 5%, or 1% (by dry weight) ofchemical precursors or non-protein-of-interest chemicals.

The term “fragment” refers to a portion of a polypeptide sequence of theinvention. “Fragments” or “biologically active portions” includepolypeptides comprising a sufficient number of contiguous amino acidresidues to retain the biological activity, i.e., have pesticidalactivity. Fragments of the pesticidal proteins include those that areshorter than the full-length sequences, either due to the use of analternate downstream start site, or due to processing that produces ashorter protein having pesticidal activity. Processing may occur in theorganism the protein is expressed in, or in the pest after ingestion ofthe protein. Examples of fragments of the proteins can be found inTable 1. A biologically active portion of a pesticidal protein can be apolypeptide that is, for example, 10, 25, 50, 100, 150, 200, 250 or moreamino acids in length of any one of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44,45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62,63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98,99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112,113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126,127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140,141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154,155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168,169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196,197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210,211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224,225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238,239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252,253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266,267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280,281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294,295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308,309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322,323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336,337, 338, 339, 340, and/or 341. Such biologically active portions can beprepared by recombinant techniques and evaluated for pesticidalactivity. As used here, a fragment comprises at least 8 contiguous aminoacids of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51,52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104,105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118,119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132,133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146,147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160,161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174,175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188,189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202,203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216,217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230,231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244,245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258,259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272,273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286,287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300,301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314,315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328,329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, and/or 341.

Bacterial genes, including those encoding the pesticidal proteinsdisclosed herein, quite often possess multiple methionine initiationcodons in proximity to the start of the open reading frame. Often,translation initiation at one or more of these start codons will lead togeneration of a functional protein. These start codons can include ATGcodons. However, bacteria such as Bacillus sp. also recognize the codonGTG as a start codon, and proteins that initiate translation at GTGcodons contain a methionine at the first amino acid. On rare occasions,translation in bacterial systems can initiate at a TTG codon, though inthis event the TTG encodes a methionine. Furthermore, it is not oftendetermined a priori which of these codons are used naturally in thebacterium. Thus, it is understood that use of one of the alternatemethionine codons may also lead to generation of pesticidal proteins.These pesticidal proteins are encompassed in the present invention andmay be used in the methods disclosed herein. It will be understood that,when expressed in plants, it will be necessary to alter the alternatestart codon to ATG for proper translation.

In various embodiments the pesticidal proteins provided herein includeamino acid sequences deduced from the full-length nucleotide sequencesand amino acid sequences that are shorter than the full-length sequencesdue to the use of an alternate downstream start site. Thus, thenucleotide sequence of the invention and/or vectors, host cells, andplants comprising the nucleotide sequence of the invention (and methodsof making and using the nucleotide sequence of the invention) maycomprise a nucleotide sequence encoding an alternate start site.

It is recognized that modifications may be made to the pesticidalpolypeptides provided herein creating variant proteins. Changes designedby man may be introduced through the application of site-directedmutagenesis techniques. Alternatively, native, as yet-unknown or as yetunidentified polynucleotides and/or polypeptides structurally and/orfunctionally-related to the sequences disclosed herein may also beidentified that fall within the scope of the present invention.Conservative amino acid substitutions may be made in nonconservedregions that do not alter the function of the pesticidal proteins.Alternatively, modifications may be made that improve the activity ofthe toxin. Modification of Cry toxins by domain III swapping hasresulted in some cases in hybrid toxins with improved toxicities againstcertain insect species. Thus, domain III swapping could be an effectivestrategy to improve toxicity of Cry toxins or to create novel hybridtoxins with toxicity against pests that show no susceptibility to theparental Cry toxins. Site-directed mutagenesis of domain II loopsequences may result in new toxins with increased insecticidal activity.Domain II loop regions are key binding regions of initial Cry toxinsthat are suitable targets for the mutagenesis and selection of Crytoxins with improved insecticidal properties. Domain I of the Cry toxinmay be modified to introduce protease cleavage sites to improve activityagainst certain pests. Strategies for shuffling the three differentdomains among large numbers of cry genes and high throughput outputbioassay screening methods may provide novel Cry toxins with improved ornovel toxicities.

As indicated, fragments and variants of the polypeptides disclosedherein will retain pesticidal activity. Pesticidal activity comprisesthe ability of the composition to achieve an observable effectdiminishing the occurrence or an activity of the target pest, includingfor example, bringing about death of at least one pest, or a noticeablereduction in pest growth, feeding, or normal physiological development.Such decreases in numbers, pest growth, feeding or normal developmentcan comprise any statistically significant decrease, including, forexample a decrease of about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%,50%, 55%, 60%, 65%, 70%, 75%, 85%, 90%, 95% or greater. It is recognizedthat the pesticidal activity may be different or improved relative tothe activity of the native protein, or it may be unchanged, so long aspesticidal activity is retained. Methods for measuring pesticidalactivity are well known in the art. See, for example, Czapla and Lang(1990) J. Econ. Entomol. 83:2480-2485; Andrews et al. (1988) Biochem. J.252:199-206; Marrone et al. (1985) J. of Economic Entomology 78:290-293;and U.S. Pat. No. 5,743,477, all of which are herein incorporated byreference in their entirety.

Polypeptide variants of this disclosure include polypeptides having anamino acid sequence that is at least about 60%, about 65%, about 70%,about 75%, about 80%, about 85%, about 86%, about 87%, about 88%, about89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%,about 96%, about 97%, about 98% or about 99% identical to the amino acidsequence of any of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30,31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48,49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66,67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84,85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101,102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115,116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129,130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143,144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157,158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171,172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185,186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199,200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213,214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227,228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241,242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255,256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269,270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283,284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297,298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311,312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325,326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339,340, and/or 341 and retain pesticidal activity. Note, Table 1 providesnon-limiting examples of variant polypeptides (and polynucleotideencoding the same) for each of SEQ ID NOS: 1-341. A biologically activevariant of a pesticidal polypeptide of the invention may differ by asfew as about 1-15 amino acid residues, as few as about 1-10, such asabout 6-10, as few as 5, as few as 4, as few as 3, as few as 2, or asfew as 1 amino acid residue. In specific embodiments, the polypeptidescan comprise an N′-terminal or a C′-terminal truncation, which cancomprise at least a deletion of 10, 15, 20, 25, 30, 35, 40, 45, 50 aminoacids or more from either the N′ or C′ terminal end of the polypeptide.

Table 2 provides protein domains found in SEQ ID NOs: 1-341 based onPFAM data. Both the domain description and the positions within a givenSEQ ID NO are provided in Table 2. In specific embodiments, the activevariant comprising any one of SEQ ID NOs: 1-341 can comprise at least70%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequenceidentity to any one of SEQ ID NOs: 1-341 and further comprises at leastone of the conserved domain set forth in Table 2.

TABLE 2 PFAM summary Modification Domain Domain Position APG ID Seq IDType PFAM Domain Description Start Stop APG00038 Seq ID 1 PF12495Vip3A_N 10 185 APG00138 Seq ID 2 PF01338 Bac_thur_toxin 131 324 APG00139Seq ID 3 PF05791 Bacillus_HBL 45 231 APG00143 Seq ID 4 PF03318 ETX_MTX222 222 APG00154 Seq ID 5 PF03318 ETX_MTX2 118 336 APG00156 Seq ID 6PF03318 ETX_MTX2 65 286 APG00157 Seq ID 7 PF14200 RicinB_lectin_2 50 150PF05431 Toxin_10 156 353 APG00158 Seq ID 8 PF03318 ETX_MTX2 89 304APG00159 Seq ID 9 PF03318 ETX_MTX2 18 273 APG00160 Seq ID 10 PF05791Bacillus_HBL 38 218 APG00165 Seq ID 11 PF03945 Endotoxin_N 122 391PF03944 Endotoxin_C 658 797 APG00165 Seq ID 12 3′ Truncation PF03945Endotoxin_N 122 391 modified PF03944 Endotoxin_C 658 796 APG00170 Seq ID13 PF03318 ETX_MTX2 134 331 APG00170 Seq ID 14 Signal PF03318 ETX_MTX280 280 modified peptide removed APG00171 Seq ID 15 PF03318 ETX_MTX2 149355 PF09259 Fve 383 471 APG00181 Seq ID 16 PF12495 Vip3A_N 10 185APG00182 Seq ID 17 PF03945 Endotoxin_N 87 317 PF03944 Endotoxin_C 513647 APG00182 Seq ID 18 3′ Truncation PF03945 Endotoxin_N 87 317 modifiedPF03944 Endotoxin_C 513 646 APG00184 Seq ID 19 PF01473 CW_binding_1 176197 APG00187 Seq ID 20 PF03318 ETX_MTX2 33 296 APG00187 Seq ID 21 SignalPF03318 ETX_MTX2 12 274 modified peptide removed APG00193 Seq ID 22PF06101 DUF946 33 208 PF05431 Toxin_10 242 426 APG00197 Seq ID 23PF05791 Bacillus_HBL 38 218 APG00217 Seq ID 24 PF03945 Endotoxin_N 70335 PF03944 Endotoxin_C 556 695 APG00217 Seq ID 25 3′ Truncation PF03945Endotoxin_N 70 335 modified PF03944 Endotoxin_C 556 694 APG00218 Seq ID26 PF03318 ETX_MTX2 39 293 APG00220 Seq ID 27 PF03318 ETX_MTX2 142 354APG00224 Seq ID 28 PF03318 ETX_MTX2 62 330 APG00225 Seq ID 29 PF05791Bacillus_HBL 50 233 APG00226 Seq ID 30 PF14200 RicinB_lectin_2 43 147PF05431 Toxin_10 154 347 APG00228 Seq ID 31 PF03318 ETX_MTX2 100 353APG00236 Seq ID 32 PF03318 ETX_MTX2 47 252 APG00244 Seq ID 33 PF03318ETX_MTX2 33 298 APG00245 Seq ID 34 PF03318 ETX_MTX2 40 293 APG00246 SeqID 35 PF03318 ETX_MTX2 114 281 APG00247 Seq ID 36 PF00652 Ricin_B_lectin65 158 PF05431 Toxin_10 224 416 APG00250 Seq ID 37 PF03945 Endotoxin_N72 282 PF01473 CW_binding_1 297 311 PF01473 CW_binding_1 380 395 PF01473CW_binding_1 434 448 APG00252 Seq ID 38 PF03318 ETX_MTX2 33 255 APG00253Seq ID 39 PF03318 ETX_MTX2 104 234 APG00254 Seq ID 40 PF03318 ETX_MTX2 1195 APG00256 Seq ID 41 PF05431 Toxin_10 156 359 APG00257 Seq ID 42PF14200 RicinB_lectin_2 43 145 PF05431 Toxin_10 151 347 APG00259 Seq ID43 PF05791 Bacillus_HBL 39 218 APG00260 Seq ID 44 PF03318 ETX_MTX2 33258 APG00268 Seq ID 45 PF03318 ETX_MTX2 127 311 APG00269 Seq ID 46PF03318 ETX_MTX2 128 357 APG00276 Seq ID 47 PF03318 ETX_MTX2 19 304APG00277 Seq ID 48 PF03318 ETX_MTX2 33 252 APG00281 Seq ID 49 PF03318ETX_MTX2 154 293 APG00283 Seq ID 50 PF03318 ETX_MTX2 59 322 APG00293 SeqID 51 PF03318 ETX_MTX2 121 306 APG00296 Seq ID 52 PF03945 Endotoxin_N 44296 PF01473 CW_binding_1 366 379 PF01473 CW_binding_1 387 403 APG00300Seq ID 53 PF05431 Toxin_10 203 396 APG00303 Seq ID 54 PF03318 ETX_MTX2105 319 APG00304 Seq ID 55 PF03318 ETX_MTX2 83 340 APG00309 Seq ID 56PF03318 ETX_MTX2 84 324 APG00309 Seq ID 57 Signal PF03318 ETX_MTX2 29294 modified peptide removed APG00321 Seq ID 58 PF03945 Endotoxin_N 70275 APG00322 Seq ID 59 PF03318 ETX_MTX2 126 251 APG00323 Seq ID 60PF03318 ETX_MTX2 55 322 APG00324 Seq ID 61 PF03318 ETX_MTX2 58 362APG00329 Seq ID 62 PF03945 Endotoxin_N 126 330 APG00330 Seq ID 63PF03945 Endotoxin_N 81 280 APG00332 Seq ID 64 PF03318 ETX_MTX2 28 266APG00339 Seq ID 65 PF03318 ETX_MTX2 46 245 APG00341 Seq ID 66 PF03318ETX_MTX2 345 495 APG00354 Seq ID 67 PF03318 ETX_MTX2 10 214 APG00359 SeqID 68 PF03318 ETX_MTX2 137 248 APG00360 Seq ID 69 PF03318 ETX_MTX2 18267 APG00359 Seq ID 70 Signal PF03318 ETX_MTX2 108 219 modified peptideremoved APG00367 Seq ID 71 PF03945 Endotoxin_N 1 174 APG00371 Seq ID 72PF03318 ETX_MTX2 108 262 APG00410 Seq ID 73 Signal PF03318 ETX_MTX2 20284 modified peptide removed APG00382 Seq ID 74 PF03318 ETX_MTX2 19 273APG00390 Seq ID 75 PF03318 ETX_MTX2 192 365 APG00393 Seq ID 76 PF03318ETX_MTX2 123 363 APG00402 Seq ID 77 PF03318 ETX_MTX2 62 273 APG00403 SeqID 78 PF05431 Toxin_10 154 358 APG00410 Seq ID 79 PF03318 ETX_MTX2 74314 APG00411 Seq ID 80 PF03318 ETX_MTX2 60 341 APG00414 Seq ID 81PF03318 ETX_MTX2 117 351 APG00416 Seq ID 82 PF03318 ETX_MTX2 77 305APG00417 Seq ID 83 PF03318 ETX_MTX2 67 298 APG00418 Seq ID 84 PF03318ETX_MTX2 92 226 APG00420 Seq ID 85 PF03318 ETX_MTX2 53 324 APG00421 SeqID 86 PF03318 ETX_MTX2 119 351 APG00425 Seq ID 87 PF03318 ETX_MTX2 67312 APG00426 Seq ID 88 PF03318 ETX_MTX2 123 356 APG00428 Seq ID 89PF03318 ETX_MTX2 130 335 APG00432 Seq ID 90 PF05791 Bacillus_HBL 50 207APG00441 Seq ID 91 PF03318 ETX_MTX2 19 236 APG00443 Seq ID 92 no PFAMdomains APG00444 Seq ID 93 PF03318 ETX_MTX2 20 214 APG00451 Seq ID 94PF10664 NdhM 28 121 PF03318 ETX_MTX2 93 319 APG00453 Seq ID 95 PF03318ETX_MTX2 17 251 APG00455 Seq ID 96 PF03945 Endotoxin_N 118 308 APG00455Seq ID 97 Alternate PF03945 Endotoxin_N 38 228 modified start APG00457Seq ID 98 PF03945 Endotoxin_N 87 288 PF01473 CW_binding_1 348 365PF01473 CW_binding_1 368 382 PF01473 CW_binding_1 404 420 PF14200RicinB_lectin_2 461 566 APG00477 Seq ID 99 PF03318 ETX_MTX2 87 325APG00477 Seq ID 100 Signal PF03318 ETX_MTX2 53 291 modified peptideremoved APG00479 Seq ID 101 PF03318 ETX_MTX2 80 307 APG00480 Seq ID 102PF03945 Endotoxin_N 41 282 APG00481 Seq ID 103 PF03318 ETX_MTX2 126 266APG00483 Seq ID 104 PF03318 ETX_MTX2 86 352 APG00484 Seq ID 105 PF03318ETX_MTX2 21 273 APG00485 Seq ID 106 no PFAM domains APG00486 Seq ID 107PF03318 ETX_MTX2 102 337 APG00488 Seq ID 108 PF03318 ETX_MTX2 157 394APG00488 Seq ID 109 Signal PF03318 ETX_MTX2 124 361 modified peptideremoved APG00496 Seq ID 110 PF03945 Endotoxin_N 56 301 APG00508 Seq ID111 PF03945 Endotoxin_N 54 263 APG00508 Seq ID 112 3′ Truncation PF03945Endotoxin_N 54 264 modified APG00509 Seq ID 113 PF03318 ETX_MTX2 77 298APG00510 Seq ID 114 PF03945 Endotoxin_N 54 255 APG00519 Seq ID 115 noPFAM domains APG00522 Seq ID 116 PF03318 ETX_MTX2 46 299 APG00522 Seq ID117 Signal PF03318 ETX_MTX2 32 272 modified peptide removed APG00535 SeqID 118 PF03318 ETX_MTX2 40 275 APG00541 Seq ID 119 PF03318 ETX_MTX2 71298 APG00542 Seq ID 120 PF03318 ETX_MTX2 127 360 APG00542 Seq ID 121Signal PF03318 ETX_MTX2 64 296 modified peptide removed APG00545 Seq ID122 PF03318 ETX_MTX2 38 319 APG00546 Seq ID 123 PF03318 ETX_MTX2 301 527APG00553 Seq ID 124 PF01473 CW_binding_1 384 399 APG00560 Seq ID 125PF03318 ETX_MTX2 127 351 APG00562 Seq ID 126 PF03318 ETX_MTX2 75 265APG00563 Seq ID 127 PF03318 ETX_MTX2 68 305 APG00564 Seq ID 128 PF03945Endotoxin_N 122 321 APG00569 Seq ID 129 PF03318 ETX_MTX2 74 306 APG00570Seq ID 130 PF03318 ETX_MTX2 1 101 APG00579 Seq ID 131 PF03318 ETX_MTX2136 335 APG00580 Seq ID 132 PF03318 ETX_MTX2 87 316 APG00581 Seq ID 133PF03945 Endotoxin_N 68 327 PF00555 Endotoxin_M 332 532 PF03944Endotoxin_C 542 683 APG00581 Seq ID 134 3′ Truncation PF03945Endotoxin_N 68 327 modified PF00555 Endotoxin_M 332 532 PF03944Endotoxin_C 542 682 APG00586 Seq ID 135 PF03318 ETX_MTX2 92 324 APG00593Seq ID 136 PF03318 ETX_MTX2 123 351 APG00594 Seq ID 137 PF03318 ETX_MTX2115 337 APG00595 Seq ID 138 PF03945 Endotoxin_N 98 341 PF03944Endotoxin_C 524 658 APG00597 Seq ID 139 PF03318 ETX_MTX2 113 262APG00598 Seq ID 140 PF03318 ETX_MTX2 11 278 APG00601 Seq ID 141 PF03318ETX_MTX2 42 256 APG00613 Seq ID 142 PF03945 Endotoxin_N 68 290 PF00555Endotoxin_M 295 504 PF03944 Endotoxin_C 514 656 APG00613 Seq ID 143 3′Truncation PF03945 Endotoxin_N 68 290 modified PF00555 Endotoxin_M 295504 PF03944 Endotoxin_C 514 655 APG00614 Seq ID 144 no PFAM domainsAPG00614 Seq ID 145 PF07029 CryBP1 36 197 CryBP1 APG00625 Seq ID 146PF03945 Endotoxin_N 102 310 APG00628 Seq ID 147 PF03318 ETX_MTX2 117 360APG00633 Seq ID 148 PF03318 ETX_MTX2 91 317 APG00634 Seq ID 149 PF03318ETX_MTX2 172 413 APG00636 Seq ID 150 PF03318 ETX_MTX2 157 356 APG00645Seq ID 151 PF03318 ETX_MTX2 63 307 APG00654 Seq ID 152 PF03945Endotoxin_N 84 307 PF00555 Endotoxin_M 312 530 PF03944 Endotoxin_C 540677 APG00654 Seq ID 153 3′ Truncation PF03945 Endotoxin_N 84 307modified PF00555 Endotoxin_M 312 530 PF03944 Endotoxin_C 540 676APG00665 Seq ID 154 PF03945 Endotoxin_N 77 311 PF03944 Endotoxin_C 561693 APG00667 Seq ID 155 PF03318 ETX_MTX2 45 274 APG00669 Seq ID 156PF14200 RicinB_lectin_2 53 150 PF05431 Toxin_10 156 353 APG00671 Seq ID157 PF03318 ETX_MTX2 65 307 APG00681 Seq ID 158 no PFAM domains APG00686Seq ID 159 PF03318 ETX_MTX2 73 302 APG00696 Seq ID 160 PF03318 ETX_MTX239 280 APG00704 Seq ID 161 PF03318 ETX_MTX2 110 257 APG00711 Seq ID 162PF03945 Endotoxin_N 74 314 PF03944 Endotoxin_C 539 677 APG00711 Seq ID163 3′ Truncation PF03945 Endotoxin_N 74 314 modified PF03944Endotoxin_C 539 676 APG00719 Seq ID 164 PF03318 ETX_MTX2 29 310 APG00733Seq ID 165 PF03945 Endotoxin_N 157 288 APG00755 Seq ID 166 PF03318ETX_MTX2 52 288 APG00755 Seq ID 167 Signal PF03318 ETX_MTX2 22 257modified peptide removed APG00764 Seq ID 168 PF03318 ETX_MTX2 111 311APG00780 Seq ID 169 PF06101 DUF946 1 213 PF05431 Toxin_10 216 407APG00793 Seq ID 170 PF03318 ETX_MTX2 85 322 APG00794 Seq ID 171 PF03318ETX_MTX2 74 311 APG00800 Seq ID 172 PF03945 Endotoxin_N 68 294 PF00555Endotoxin_M 299 506 PF03944 Endotoxin_C 516 647 APG00820 Seq ID 173PF03318 ETX_MTX2 70 264 APG00825 Seq ID 174 PF03318 ETX_MTX2 32 296APG00833 Seq ID 175 PF03318 ETX_MTX2 37 240 APG00834 Seq ID 176 PF03318ETX_MTX2 139 357 APG00846 Seq ID 177 PF03318 ETX_MTX2 52 327 APG00850Seq ID 178 PF03318 ETX_MTX2 35 251 APG00854 Seq ID 179 PF03318 ETX_MTX298 244 APG00860 Seq ID 180 PF03318 ETX_MTX2 75 297 APG00861 Seq ID 181PF03945 Endotoxin_N 71 292 PF00555 Endotoxin_M 297 492 PF03944Endotoxin_C 502 639 APG00861 Seq ID 182 3′ Truncation PF03945Endotoxin_N 71 292 modified PF00555 Endotoxin_M 297 492 PF03944Endotoxin_C 502 638 APG00866 Seq ID 183 PF03318 ETX_MTX2 25 277 APG00891Seq ID 184 PF03318 ETX_MTX2 140 373 APG00896 Seq ID 185 PF03318 ETX_MTX232 295 APG00897 Seq ID 186 PF03945 Endotoxin_N 61 293 PF00555Endotoxin_M 298 494 PF03944 Endotoxin_C 505 646 APG00897 Seq ID 187 3′Truncation PF03945 Endotoxin_N 61 293 modified PF00555 Endotoxin_M 298494 PF03944 Endotoxin_C 505 645 APG00899 Seq ID 188 PF03318 ETX_MTX2 5309 APG00911 Seq ID 189 PF03318 ETX_MTX2 96 247 APG00938 Seq ID 190PF03318 ETX_MTX2 74 302 APG00940 Seq ID 191 PF03318 ETX_MTX2 45 296APG00950 Seq ID 192 PF03945 Endotoxin_N 64 319 PF03944 Endotoxin_C 545686 APG00950 Seq ID 193 3′ Truncation PF03945 Endotoxin_N 64 319modified PF03944 Endotoxin_C 545 685 APG00959 Seq ID 194 PF03318ETX_MTX2 103 261 APG00966 Seq ID 195 PF03318 ETX_MTX2 51 317 APG00971Seq ID 196 PF03945 Endotoxin_N 67 290 PF00555 Endotoxin_M 295 515PF03944 Endotoxin_C 525 670 APG00971 Seq ID 197 3′ Truncation PF03945Endotoxin_N 67 290 modified PF00555 Endotoxin_M 295 515 PF03944Endotoxin_C 525 669 APG00994 Seq ID 198 PF03318 ETX_MTX2 95 345 APG00994Seq ID 199 Signal PF03318 ETX_MTX2 61 311 modified peptide removedAPG00995 Seq ID 200 PF03318 ETX_MTX2 123 351 APG00646 Seq ID 201 PF03318ETX_MTX2 126 266 APG00646 Seq ID 202 Alternate PF03318 ETX_MTX2 108 248modified start APG00646 Seq ID 203 Signal PF03318 ETX_MTX2 77 215modified peptide removed APG00138 Seq ID 204 Alternate PF01338Bac_thur_toxin 1 191 modified start APG00143 Seq ID 205 Signal PF03318ETX_MTX2 17 195 modified peptide removed APG00154 Seq ID 206 SignalPF03318 ETX_MTX2 88 307 modified peptide removed APG00156 Seq ID 207Signal PF03318 ETX_MTX2 11 232 modified peptide removed APG00157 Seq ID208 Alternate PF14200 RicinB_lectin_2 43 146 modified start PF05431Toxin_10 152 349 APG00158 Seq ID 209 Alternate PF03318 ETX_MTX2 22 239modified start APG00165 Seq ID 210 Alternate PF03945 Endotoxin_N 69 338modified start PF03944 Endotoxin_C 605 744 APG00165 Seq ID 211 AlternatePF03945 Endotoxin_N 69 338 modified start and 3′ PF03944 Endotoxin_C 605743 Truncation APG00171 Seq ID 212 Signal PF03318 ETX_MTX2 105 313modified peptide PF09259 Fve 340 428 removed APG00171 Seq ID 213 Signalpeptide PF03318 ETX_MTX2 103 316 modified removed and 3′ TruncationAPG00182 Seq ID 214 Alternate PF03945 Endotoxin_N 71 301 modified startPF03944 Endotoxin_C 497 631 APG00182 Seq ID 215 Alternate PF03945Endotoxin_N 71 301 modified start and 3′ PF03944 Endotoxin_C 497 630Truncation APG00218 Seq ID 216 Alternate PF03318 ETX_MTX2 36 290modified start APG00220 Seq ID 217 Signal PF03318 ETX_MTX2 106 320modified peptide removed APG00225 Seq ID 218 Alternate PF05791Bacillus_HBL 47 230 modified start APG00228 Seq ID 219 Alternate PF03318ETX_MTX2 97 350 modified start APG00228 Seq ID 220 Signal PF03318ETX_MTX2 69 322 modified peptide removed APG00244 Seq ID 221 SignalPF03318 ETX_MTX2 5 268 modified peptide removed APG00245 Seq ID 222Alternate PF03318 ETX_MTX2 37 290 modified start APG00246 Seq ID 223Alternate PF03318 ETX_MTX2 107 274 modified start APG00246 Seq ID 224Signal PF03318 ETX_MTX2 91 227 modified peptide removed APG00247 Seq ID225 Signal PF00652 Ricin_B_lectin 8 101 modified peptide PF05431Toxin_10 167 359 removed APG00252 Seq ID 226 Alternate PF03318 ETX_MTX231 253 modified start APG00253 Seq ID 227 Signal PF03318 ETX_MTX2 75 203modified peptide removed APG00254 Seq ID 228 Alternate PF03318 ETX_MTX21 195 modified start APG00256 Seq ID 229 Alternate PF05431 Toxin_10 125328 modified start APG00259 Seq ID 230 Alternate PF05791 Bacillus_HBL 39218 modified start APG00260 Seq ID 231 Alternate PF03318 ETX_MTX2 26 251modified start APG00268 Seq ID 232 Alternate PF03318 ETX_MTX2 108 289modified start APG00268 Seq ID 233 Signal PF03318 ETX_MTX2 77 255modified peptide removed APG00269 Seq ID 234 Alternate PF03318 ETX_MTX2122 351 modified start APG00269 Seq ID 235 Signal PF03318 ETX_MTX2 98322 modified peptide removed APG00276 Seq ID 236 Signal PF03318 ETX_MTX23 275 modified peptide removed APG00281 Seq ID 237 Signal PF03318ETX_MTX2 120 246 modified peptide removed APG00283 Seq ID 238 AlternatePF03318 ETX_MTX2 51 314 modified start APG00283 Seq ID 239 SignalPF03318 ETX_MTX2 24 287 modified peptide removed APG00293 Seq ID 240Signal PF03318 ETX_MTX2 95 290 modified peptide removed APG00296 Seq ID241 Alternate PF03945 Endotoxin_N 42 294 modified start PF01473CW_binding_1 364 377 PF01473 CW_binding_1 385 401 APG00300 Seq ID 242Signal PF05431 Toxin_10 174 367 modified peptide removed APG00303 Seq ID243 Alternate PF03318 ETX_MTX2 102 312 modified start APG00303 Seq ID244 Signal PF03318 ETX_MTX2 74 308 modified peptide removed APG00304 SeqID 245 Alternate PF03318 ETX_MTX2 74 331 modified start APG00321 Seq ID246 Alternate PF03945 Endotoxin_N 70 275 modified start APG00322 Seq ID247 Signal PF03318 ETX_MTX2 79 204 modified peptide removed APG00323 SeqID 248 Alternate PF03318 ETX_MTX2 50 317 modified start APG00324 Seq ID249 Signal PF03318 ETX_MTX2 15 320 modified peptide removed APG00329 SeqID 250 Signal PF03945 Endotoxin_N 72 276 modified peptide removedAPG00330 Seq ID 251 Alternate PF03945 Endotoxin_N 42 241 modified startAPG00330 Seq ID 252 Signal PF03945 Endotoxin_N 18 217 modified peptideremoved APG00339 Seq ID 253 Signal PF03318 ETX_MTX2 19 214 modifiedpeptide removed APG00367 Seq ID 254 Alternate PF03945 Endotoxin_N 1 174modified start APG00371 Seq ID 255 Signal PF03318 ETX_MTX2 75 256modified peptide removed APG00382 Seq ID 256 Signal PF03318 ETX_MTX2 7249 modified peptide removed APG00393 Seq ID 257 Alternate PF03318ETX_MTX2 90 345 modified start APG00393 Seq ID 258 Signal PF03318ETX_MTX2 73 314 modified peptide removed APG00403 Seq ID 259 AlternatePF05431 Toxin_10 154 358 modified start APG00403 Seq ID 260 AlternatePF05431 Toxin_10 84 288 modified start APG00411 Seq ID 261 AlternatePF03318 ETX_MTX2 37 318 modified start APG00411 Seq ID 262 SignalPF03318 ETX_MTX2 45 291 modified peptide removed APG00414 Seq ID 263Signal PF03318 ETX_MTX2 87 322 modified peptide removed APG00416 Seq ID264 Signal PF03318 ETX_MTX2 44 273 modified peptide removed APG00417 SeqID 265 Signal PF03318 ETX_MTX2 34 265 modified peptide removed APG00418Seq ID 266 Signal PF03318 ETX_MTX2 73 202 modified peptide removedAPG00420 Seq ID 267 Alternate PF03318 ETX_MTX2 48 319 modified startAPG00420 Seq ID 268 Signal PF03318 ETX_MTX2 9 275 modified peptideremoved APG00421 Seq ID 269 Signal PF03318 ETX_MTX2 90 322 modifiedpeptide removed APG00426 Seq ID 270 Signal PF03318 ETX_MTX2 94 327modified peptide removed APG00428 Seq ID 271 Signal PF03318 ETX_MTX2 115320 modified peptide removed APG00441 Seq ID 272 Signal PF03318 ETX_MTX216 209 modified peptide removed APG00443 Seq ID 273 Alternate no PFAMmodified start domains APG00444 Seq ID 274 Signal PF03318 ETX_MTX2 15188 modified peptide removed APG00451 Seq ID 275 Signal PF10664 NdhM 191 modified peptide PF03318 ETX_MTX2 62 288 removed APG00453 Seq ID 276Alternate PF03318 ETX_MTX2 15 248 modified start APG00479 Seq ID 277Signal PF03318 ETX_MTX2 54 280 modified peptide removed APG00480 Seq ID278 Alternate PF03945 Endotoxin_N 40 278 modified start APG00481 Seq ID279 Alternate PF03318 ETX_MTX2 108 249 modified start APG00481 Seq ID280 Signal PF03318 ETX_MTX2 77 216 modified peptide removed APG00483 SeqID 281 Signal PF03318 ETX_MTX2 53 324 modified peptide removed APG00484Seq ID 282 Signal PF03318 ETX_MTX2 11 249 modified peptide removedAPG00486 Seq ID 283 Signal PF03318 ETX_MTX2 73 308 modified peptideremoved APG00496 Seq ID 284 Alternate PF03945 Endotoxin_N 53 298modified start APG00509 Seq ID 285 Signal PF03318 ETX_MTX2 47 268modified peptide removed APG00510 Seq ID 286 Signal no PFAM modifiedpeptide domains removed APG00541 Seq ID 287 Signal PF03318 ETX_MTX2 40268 modified peptide removed APG00545 Seq ID 288 Signal PF03318 ETX_MTX214 290 modified peptide removed APG00560 Seq ID 289 Signal PF03318ETX_MTX2 97 322 modified peptide removed APG00562 Seq ID 290 SignalPF03318 ETX_MTX2 41 271 modified peptide removed APG00563 Seq ID 291Signal PF03318 ETX_MTX2 39 273 modified peptide removed APG00564 Seq ID292 Signal PF03945 Endotoxin_N 83 281 modified peptide removed APG00569Seq ID 293 Signal PF03318 ETX_MTX2 41 273 modified peptide removedAPG00579 Seq ID 294 Signal PF03318 ETX_MTX2 52 258 modified peptideremoved APG00580 Seq ID 295 Signal PF03318 ETX_MTX2 58 285 modifiedpeptide removed APG00586 Seq ID 296 Alternate PF03318 ETX_MTX2 87 319modified start APG00593 Seq ID 297 Signal PF03318 ETX_MTX2 94 322modified peptide removed APG00594 Seq ID 298 Signal PF03318 ETX_MTX2 85308 modified peptide removed APG00595 Seq ID 299 Alternate PF03945Endotoxin_N 1 236 modified start PF03944 Endotoxin_C 419 553 APG00595Seq ID 300 Alternate PF03945 Endotoxin_N 1 236 modified start and 3′PF03944 Endotoxin_C 419 552 Truncation APG00597 Seq ID 301 SignalPF03318 ETX_MTX2 64 211 modified peptide removed APG00601 Seq ID 302Alternate PF03318 ETX_MTX2 37 251 modified start APG00625 Seq ID 303Signal PF03945 Endotoxin_N 64 272 modified peptide removed APG00628 SeqID 304 Signal PF03318 ETX_MTX2 81 324 modified peptide removed APG00633Seq ID 305 Signal PF03318 ETX_MTX2 58 285 modified peptide removedAPG00634 Seq ID 306 Signal PF03318 ETX_MTX2 75 319 modified peptideremoved APG00636 Seq ID 307 Signal PF03318 ETX_MTX2 118 317 modifiedpeptide removed APG00654 Seq ID 308 Alternate PF03945 Endotoxin_N 72 295modified start PF00555 Endotoxin_M 300 518 PF03944 Endotoxin_C 528 665APG00665 Seq ID 309 3′ Truncation PF03945 Endotoxin_N 77 311 modifiedPF03944 Endotoxin_C 561 693 APG00667 Seq ID 310 Signal PF03318 ETX_MTX228 256 modified peptide removed APG00669 Seq ID 311 Alternate PF14200RicinB_lectin_2 2 98 modified start PF05431 Toxin_10 152 349 APG00671Seq ID 312 Signal PF03318 ETX_MTX2 36 278 modified peptide removedAPG00681 Seq ID 313 Signal PF03318 ETX_MTX2 92 342 modified peptideremoved APG00704 Seq ID 314 Signal PF03318 ETX_MTX2 80 222 modifiedpeptide removed APG00719 Seq ID 315 Alternate PF03318 ETX_MTX2 22 303modified start APG00719 Seq ID 316 Signal PF03318 ETX_MTX2 33 271modified peptide removed APG00764 Seq ID 317 Signal PF03318 ETX_MTX2 63314 modified peptide removed APG00793 Seq ID 318 Alternate PF03318ETX_MTX2 71 309 modified start APG00793 Seq ID 319 Signal PF03318ETX_MTX2 44 281 modified peptide removed APG00794 Seq ID 320 SignalPF03318 ETX_MTX2 41 278 modified peptide removed APG00800 Seq ID 321 3′Truncation PF03945 Endotoxin_N 68 294 modified PF00555 Endotoxin_M 299506 PF03944 Endotoxin_C 516 646 APG00820 Seq ID 322 Signal PF03318ETX_MTX2 8 199 modified peptide removed APG00825 Seq ID 323 SignalPF03318 ETX_MTX2 46 270 modified peptide removed APG00833 Seq ID 324Signal PF03318 ETX_MTX2 17 213 modified peptide removed APG00834 Seq ID325 Signal PF03318 ETX_MTX2 97 316 modified peptide removed APG00854 SeqID 326 Signal PF03318 ETX_MTX2 67 216 modified peptide removed APG00891Seq ID 327 Alternate PF03318 ETX_MTX2 122 355 modified start APG00891Seq ID 328 Signal PF03318 ETX_MTX2 93 326 modified peptide removedAPG00896 Seq ID 329 Signal PF03318 ETX_MTX2 4 266 modified peptideremoved APG00897 Seq ID 330 Alternate PF03945 Endotoxin_N 23 255modified start PF00555 Endotoxin_M 260 456 PF03944 Endotoxin_C 467 608APG00897 Seq ID 331 Alternate PF03945 Endotoxin_N 23 255 modified startand 3′ PF00555 Endotoxin_M 260 456 Truncation PF03944 Endotoxin_C 467607 APG00911 Seq ID 332 Signal PF03318 ETX_MTX2 68 248 modified peptideremoved APG00938 Seq ID 333 Signal PF03318 ETX_MTX2 41 269 modifiedpeptide removed APG00940 Seq ID 334 Signal PF03318 ETX_MTX2 11 259modified peptide removed APG00959 Seq ID 335 Alternate PF03318 ETX_MTX287 242 modified start APG00959 Seq ID 336 Signal PF03318 ETX_MTX2 57 297modified peptide removed APG00966 Seq ID 337 Alternate PF03318 ETX_MTX248 314 modified start APG00966 Seq ID 338 Signal PF03318 ETX_MTX2 25 290modified peptide removed APG00971 Seq ID 339 Alternate PF03945Endotoxin_N 1 199 modified start PF00555 Endotoxin_M 204 424 PF03944Endotoxin_C 434 579 APG00971 Seq ID 340 Alternate PF03945 Endotoxin_N 1199 modified start and 3′ PF00555 Endotoxin_M 204 424 Truncation PF03944Endotoxin_C 434 578 APG00995 Seq ID 341 Signal PF03318 ETX_MTX2 93 322modified peptide removed

Recombinant or synthetic nucleic acids encoding the pesticidalpolypeptides disclosed herein are also provided. Of particular interestare nucleic acid sequences that have been designed for expression in aplant of interest. That is, the nucleic acid sequence can be optimizedfor increased expression in a host plant. A pesticidal protein of theinvention can be back-translated to produce a nucleic acid comprisingcodons optimized for expression in a particular host, for example, acrop plant. In another embodiment, the polynucleotides encoding thepolypeptides provided herein may be optimized for increased expressionin the transformed plant. That is, the polynucleotides can besynthesized using plant-preferred codons for improved expression. See,for example, Campbell and Gowri (1990) Plant Physiol. 92:1-11 for adiscussion of host-preferred codon usage. Methods are available in theart for synthesizing plant-preferred genes. See, for example, U.S. Pat.Nos. 5,380,831, and 5,436,391, and Murray et al. (1989) Nucleic AcidsRes. 17:477-498, herein incorporated by reference. Expression of such acoding sequence by the transformed plant (e.g., dicot or monocot) willresult in the production of a pesticidal polypeptide and conferincreased resistance in the plant to a pest. Recombinant and syntheticnucleic acid molecules encoding the pesticidal proteins of the inventiondo not include the naturally occurring bacterial sequence encoding theprotein.

A “recombinant polynucleotide” or “recombinant nucleic acid” comprises acombination of two or more chemically linked nucleic acid segments whichare not found directly joined in nature. By “directly joined” isintended the two nucleic acid segments are immediately adjacent andjoined to one another by a chemical linkage. In specific embodiments,the recombinant polynucleotide comprises a polynucleotide of interest ora variant or fragment thereof such that an additional chemically linkednucleic acid segment is located either 5′, 3′ or internal to thepolynucleotide of interest. Alternatively, the chemically-linked nucleicacid segment of the recombinant polynucleotide can be formed by deletionof a sequence. The additional chemically linked nucleic acid segment orthe sequence deleted to join the linked nucleic acid segments can be ofany length, including for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20or greater nucleotides. Various methods for making such recombinantpolynucleotides include chemical synthesis or by the manipulation ofisolated segments of polynucleotides by genetic engineering techniques.In specific embodiments, the recombinant polynucleotide can comprise arecombinant DNA sequence or a recombinant RNA sequence. A “fragment of arecombinant polynucleotide or nucleic acid” comprises at least one of acombination of two or more chemically linked amino acid segments whichare not found directly joined in nature.

Fragments of a polynucleotide (RNA or DNA) may encode protein fragmentsthat retain activity. In specific embodiments, a fragment of arecombinant polynucleotide or a recombinant polynucleotide constructcomprises at least one junction of the two or more chemically linked oroperably linked nucleic acid segments which are not found directlyjoined in nature. A fragment of a polynucleotide that encodes abiologically active portion of a polypeptide that retains pesticidalactivity will encode at least 25, 30, 40, 50, 60, 70, 75, 80, 90, 100,110, 120, 125, 130, 140, 150, 160, 170, 175, 180, contiguous aminoacids, or up to the total number of amino acids present in a full-lengthpolypeptide as set forth in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28,29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46,47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64,65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100,101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114,115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128,129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142,143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156,157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170,171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184,185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198,199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226,227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240,241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254,255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268,269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282,283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296,297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310,311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324,325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338,339, 340, and/or 341. In specific embodiments, such polypeptidefragments are active fragments, and in still other embodiments, thepolypeptide fragment comprises a recombinant polypeptide fragment. Asused herein, a fragment of a recombinant polypeptide comprises at leastone of a combination a two or more chemically linked amino acid segmentswhich are not found directly joined in nature.

By “variants” is intended to mean substantially similar sequences. Forpolynucleotides, a variant comprises a deletion and/or addition of oneor more nucleotides at one or more internal sites within the nativepolynucleotide and/or a substitution of one or more nucleotides at oneor more sites in the native polynucleotide. As used herein, a “native”polynucleotide or polypeptide comprises a naturally occurring nucleotidesequence or amino acid sequence, respectively.

Variants of a particular polynucleotide of the invention (i.e., thereference polynucleotide) can also be evaluated by comparison of thepercent sequence identity between the polypeptide encoded by a variantpolynucleotide and the polypeptide encoded by the referencepolynucleotide. Thus, for example, an isolated polynucleotide thatencodes a polypeptide with a given percent sequence identity to thepolypeptide of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32,33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50,51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68,69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86,87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103,104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117,118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131,132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145,146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159,160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173,174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187,188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201,202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215,216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229,230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243,244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257,258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271,272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285,286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299,300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313,314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327,328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, and/or341 are disclosed. Percent sequence identity between any twopolypeptides can be calculated using sequence alignment programs andparameters described elsewhere herein. Where any given pair ofpolynucleotides of the invention is evaluated by comparison of thepercent sequence identity shared by the two polypeptides they encode,the percent sequence identity between the two encoded polypeptides is atleast about 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or moresequence identity to SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30,31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48,49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66,67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84,85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101,102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115,116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129,130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143,144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157,158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171,172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185,186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199,200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213,214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227,228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241,242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255,256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269,270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283,284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297,298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311,312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325,326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339,340, and/or 341. In other embodiments, the variant of the polynucleotideprovided herein differs from the native sequence by at least 1, 2, 3, 4,5, 6, 7, 8, 9, 10 or more nucleotides.

Variant polynucleotide and proteins also encompass sequences andproteins derived from a mutagenic and recombinogenic procedure such asDNA shuffling. With such a procedure, one or more different pesticidalprotein disclosed herein (SEQ ID NO: 1-341) is manipulated to create anew pesticidal protein possessing the desired properties. In thismanner, libraries of recombinant polynucleotides are generated from apopulation of related sequence polynucleotides comprising sequenceregions that have substantial sequence identity and can be homologouslyrecombined in vitro or in vivo. For example, using this approach,sequence motifs encoding a domain of interest may be shuffled betweenthe pesticidal sequences provided herein and other known pesticidalgenes to obtain a new gene coding for a protein with an improvedproperty of interest, such as an increased K_(m) in the case of anenzyme. Strategies for such DNA shuffling are known in the art. See, forexample, Stemmer (1994) Proc. Natl. Acad. Sci. USA 91:10747-10751;Stemmer (1994) Nature 370:389-391; Crameri et al. (1997) Nature Biotech.15:436-438; Moore et al. (1997) J. Mol. Biol. 272:336-347; Zhang et al.(1997) Proc. Natl. Acad. Sci. USA 94:4504-4509; Crameri et al. (1998)Nature 391:288-291; and U.S. Pat. Nos. 5,605,793 and 5,837,458. A“shuffled” nucleic acid is a nucleic acid produced by a shufflingprocedure such as any shuffling procedure set forth herein. Shufflednucleic acids are produced by recombining (physically or virtually) twoor more nucleic acids (or character strings), for example in anartificial, and optionally recursive, fashion. Generally, one or morescreening steps are used in shuffling processes to identify nucleicacids of interest; this screening step can be performed before or afterany recombination step. In some (but not all) shuffling embodiments, itis desirable to perform multiple rounds of recombination prior toselection to increase the diversity of the pool to be screened. Theoverall process of recombination and selection are optionally repeatedrecursively. Depending on context, shuffling can refer to an overallprocess of recombination and selection, or, alternately, can simplyrefer to the recombinational portions of the overall process.

In one embodiment, a method of obtaining a polynucleotide that encodesan improved polypeptide comprising pesticidal activity is provided,wherein the improved polypeptide has at least one improved property overany one of SEQ ID NOS: 1-341. Such methods can comprise (a) recombininga plurality of parental polynucleotides to produce a library ofrecombinant polynucleotides encoding recombinant pesticidalpolypeptides; (b) screening the library to identify a recombinantpolynucleotide that encodes an improved recombinant pesticidalpolypeptide that has an enhanced property improved over the parentalpolynucleotide; (c) recovering the recombinant polynucleotide thatencodes the improved recombinant pesticidal polypeptide identified in(b); and, (d) repeating steps (a), (b) and (c) using the recombinantpolynucleotide recovered in step (c) as one of the plurality of parentalpolynucleotides in repeated step (a).

iii. Sequence Comparisons

As used herein, the term “identity” or “percent identity” when used withrespect to a particular pair of aligned amino acid sequences, refers tothe percent amino acid sequence identity that is obtained by countingthe number of identical matches in the alignment and dividing suchnumber of identical matches by the length of the aligned sequences. Asused herein, the term “similarity” or “percent similarity” when usedwith respect to a particular pair of aligned amino acid sequences,refers to the sum of the scores that are obtained from a scoring matrixfor each amino acid pair in the alignment divided by the length of thealigned sequences.

Unless otherwise stated, identity and similarity will be calculated bythe Needleman-Wunsch global alignment and scoring algorithms (Needlemanand Wunsch (1970) J. Mol. Biol. 48(3):443-453) as implemented by the“needle” program, distributed as part of the EMBOSS software package(Rice, P. Longden, I. and Bleasby, A., EMBOSS: The European MolecularBiology Open Software Suite, 2000, Trends in Genetics 16(6): 276-277,versions 6.3.1 available from EMBnet at embnet.org/resource/emboss andemboss.sourceforge.net, among other sources) using default gap penaltiesand scoring matrices (EBLOSUM62 for protein and EDNAFULL for DNA).Equivalent programs may also be used. By “equivalent program” isintended any sequence comparison program that, for any two sequences inquestion, generates an alignment having identical nucleotide residuematches and an identical percent sequence identity when compared to thecorresponding alignment generated by needle from EMBOSS version 6.3.1.

Additional mathematical algorithms are known in the art and can beutilized for the comparison of two sequences. See, for example, thealgorithm of Karlin and Altschul (1990) Proc. Natl. Acad. Scil. USA87:2264, modified as in Karlin and Altschul (1993) Proc. Natl. Acad.Sci. USA 90:5873-5877. Such an algorithm is incorporated into the BLASTprograms of Altschul et al. (1990) J. Mol. Biol. 215:403. BLASTnucleotide searches can be performed with the BLASTN program (nucleotidequery searched against nucleotide sequences) to obtain nucleotidesequences homologous to pesticidal-like nucleic acid molecules of theinvention, or with the BLASTX program (translated nucleotide querysearched against protein sequences) to obtain protein sequenceshomologous to pesticidal nucleic acid molecules of the invention. BLASTprotein searches can be performed with the BLASTP program (protein querysearched against protein sequences) to obtain amino acid sequenceshomologous to pesticidal protein molecules of the invention, or with theTBLASTN program (protein query searched against translated nucleotidesequences) to obtain nucleotide sequences homologous to pesticidalprotein molecules of the invention. To obtain gapped alignments forcomparison purposes, Gapped BLAST (in BLAST 2.0) can be utilized asdescribed in Altschul et al. (1997) Nucleic Acids Res. 25:3389.Alternatively, PSI-Blast can be used to perform an iterated search thatdetects distant relationships between molecules. See Altschul et al.(1997) supra. When utilizing BLAST, Gapped BLAST, and PSI-Blastprograms, the default parameters of the respective programs (e.g.,BLASTX and BLASTN) can be used. Alignment may also be performed manuallyby inspection.

Two sequences are “optimally aligned” when they are aligned forsimilarity scoring using a defined amino acid substitution matrix (e.g.,BLOSUM62), gap existence penalty and gap extension penalty so as toarrive at the highest score possible for that pair of sequences. Aminoacid substitution matrices and their use in quantifying the similaritybetween two sequences are well-known in the art and described, e.g., inDayhoff et al. (1978) “A model of evolutionary change in proteins.” In“Atlas of Protein Sequence and Structure,” Vol. 5, Suppl. 3 (ed. M. O.Dayhoff), pp. 345-352. Natl. Biomed. Res. Found., Washington, D.C., andHenikoff et al. (1992) Proc. Natl. Acad. Sci. USA 89:10915-10919. TheBLOSUM62 matrix is often used as a default scoring substitution matrixin sequence alignment protocols. The gap existence penalty is imposedfor the introduction of a single amino acid gap in one of the alignedsequences, and the gap extension penalty is imposed for each additionalempty amino acid position inserted into an already opened gap. Thealignment is defined by the amino acids positions of each sequence atwhich the alignment begins and ends, and optionally by the insertion ofa gap or multiple gaps in one or both sequences, so as to arrive at thehighest possible score. While optimal alignment and scoring can beaccomplished manually, the process is facilitated by the use of acomputer-implemented alignment algorithm, e.g., gapped BLAST 2.0,described in Altschul et al. (1997) Nucleic Acids Res. 25:3389-3402, andmade available to the public at the National Center for BiotechnologyInformation Website (www.ncbi.nlm.nih.gov). Optimal alignments,including multiple alignments, can be prepared using, e.g., PSI-BLAST,available through www.ncbi.nlm.nih.gov and described by Altschul et al.(1997) Nucleic Acids Res. 25:3389-3402.

With respect to an amino acid sequence that is optimally aligned with areference sequence, an amino acid residue “corresponds to” the positionin the reference sequence with which the residue is paired in thealignment. The “position” is denoted by a number that sequentiallyidentifies each amino acid in the reference sequence based on itsposition relative to the N-terminus. For example, in SEQ ID NO: 1position 1 is M, position 2 is V, position 3 is Q, etc. When a testsequence is optimally aligned with SEQ ID NO: 1, a residue in the testsequence that aligns with the Q at position 3 is said to “correspond toposition 3” of SEQ ID NO: 1. Owing to deletions, insertion, truncations,fusions, etc., that must be taken into account when determining anoptimal alignment, in general the amino acid residue number in a testsequence as determined by simply counting from the N-terminal will notnecessarily be the same as the number of its corresponding position inthe reference sequence. For example, in a case where there is a deletionin an aligned test sequence, there will be no amino acid thatcorresponds to a position in the reference sequence at the site ofdeletion. Where there is an insertion in an aligned reference sequence,that insertion will not correspond to any amino acid position in thereference sequence. In the case of truncations or fusions there can bestretches of amino acids in either the reference or aligned sequencethat do not correspond to any amino acid in the corresponding sequence.

iv. Antibodies

Antibodies to the polypeptides of the present invention, or to variantsor fragments thereof, are also encompassed. Methods for producingantibodies are well known in the art (see, for example, Harlow and Lane(1988) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory,Cold Spring Harbor, N.Y.; and U.S. Pat. No. 4,196,265). These antibodiescan be used in kits for the detection and isolation of toxinpolypeptides. Thus, this disclosure provides kits comprising antibodiesthat specifically bind to the polypeptides described herein, including,for example, polypeptides having the sequence of SEQ ID NOs: 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59,60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77,78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110,111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124,125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138,139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152,153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166,167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180,181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194,195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208,209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222,223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236,237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250,251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264,265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278,279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292,293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306,307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320,321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334,335, 336, 337, 338, 339, 340, and/or 341.

II. Pests

The compositions and methods provided herein are useful against avariety of pests. “Pests” includes but is not limited to, insects,fungi, bacteria, nematodes, acarids, is protozoan pathogens,animal-parasitic liver flukes, and the like. Pests of particularinterest are insect pests, particularly insect pests that causesignificant damage to agricultural plants. Insect pests include insectsselected from the orders Coleoptera, Diptera, Hymenoptera, Lepidoptera,Mallophaga, Homoptera, Hemiptera, Orthroptera, Thysanoptera, Dermaptera,Isoptera, Anoplura, Siphonaptera, Trichoptera, or nematodes. Innon-limiting embodiments, the insect pest comprises Western cornrootworm, Diabrotica virgifera virgifera; Fall armyworm, Spodopterafrupperda; Colorado potato beetle, Leptinotarsa decemlineata; Cornearworm, Helicoverpa zea (in North America same species attacks cottonand called cotton bollworm); European corn borer, Ostrinia nubilalis;Black cutworm, Agrotis ipsilon; Diamondback moth, Plutella xylostella;Velvetbean caterpillar, Anticarsia gemmatalis; Southwestern corn borer,Diatraea grandiosella; Cotton bollworm, Helicoverpa armigera (foundother than USA in rest of the world); Southern green stinkbug, Nezaraviridula; Green stinkbug, Chinavia halaris; Brown marmorated stinkbug,Halyomorpha halys; and Brown stinbug, Euschistus servus Euschistus heros(Neotropical brown stink bug OR soy stink bug); Piezodorus guildinii(red-banded stink bug); Dichelops melacanthus (no common name) and/orDichelops furcatus (no common name); an aphid, such as a soybean aphid.In other embodiments, the pest comprises a nematode including, but notlimited to, Meloidogyne hapla (Northern root-knot nematode); Meloidogyneenterolobii, Meloidogyne arenaria (peanut root-knot nematode); andMeloidogyne javanica.

The term “insect pests” as used herein refers to insects and othersimilar pests such as, for example, those of the order Acari including,but not limited to, mites and ticks. Insect pests of the presentinvention include, but are not limited to, insects of the orderLepidoptera, e.g., Achoroia grisella, Acleris gloverana, Aclerisvariana, Adoxophyes orana, Agrotis ipsilon, Alabama argillacea,Alsophila pometaria, Amyelois transitella, Anagasta kuehniella, Anarsialineatella, Anisota senatoria, Antheraea pernyi, Anticarsia gemmatalis,Archips sp., Argyrotaenia sp., Athetis mindara, Bombyx mori, Bucculatrixthurberiella, Cadra cautella, Choristoneura sp., Cochylls hospes, Coliaseurytheme, Corcyra cephalonica, Cydia latiferreanus, Cydia pomonella,Datana integerrima, Dendrolimus sibericus, Desmiafeneralis, Diaphaniahyalinata, Diaphania nitidalis, Diatraea grandiosella, Diatraeasaccharalis, Ennomos subsignaria, Eoreuma loftini, Esphestia elutella,Erannis tilaria, Estigmene acrea, Eulia salubricola, Eupocoelliaambiguella, Eupoecilia ambiguella, Euproctis chrysorrhoea, Euxoamessoria, Galleria mellonella, Grapholita molesta, Harrisina americana,Helicoverpa subflexa, Helicoverpa zea, Heliothis virescens, Hemileucaoliviae, Homoeosoma electellum, Hyphantia cunea, Keiferialycopersicella, Lambdina fiscellaria fiscellaria, Lambdina fiscellarialugubrosa, Leucoma salicis, Lobesia botrana, Loxostege sticticalis,Lymantria dispar, Macalla thyrisalis, Malacosoma sp., Mamestrabrassicae, Mamestra configurata, Manduca quinquemaculata, Manduca sexta,Maruca testulalis, Melanchra picta, Operophtera brumata, Orgyia sp.,Ostrinia nubilalis, Paleacrita vernata, Papilio cresphontes,Pectinophora gossypiella, Phryganidia californica, Phyllonorycterblancardella, Pieris napi, Pieris rapae, Plathypena scabra, Platynotaflouendana, Platynota stultana, Platyptilia carduidactyla, Plodiainterpunctella, Plutella xylostella, Pontia protodice, Pseudaletiaunipuncta, Pseudoplasia includens, Sabulodes aegrotata, Schizuraconcinna, Sitotroga cerealella, Spilonta ocellana, Spodoptera sp.,Thaurnstopoea pityocampa, Tinsola bisselliella, Trichoplusia hi, Udearubigalis, Xylomyges curiails, and Yponomeuta padella.

Insect pests also include insects selected from the orders Diptera,Hymenoptera, Lepidoptera, Mallophaga, Homoptera, Hemiptera, Orthroptera,Thysanoptera, Dermaptera, Isoptera, Anoplura, Siphonaptera, Trichoptera,and Coleoptera.

Insect pests of the invention for the major crops include, but are notlimited to: Maize: Ostrinia nubilalis, European corn borer; Agrotisipsilon, black cutworm; Helicoverpa zeae, corn earworm; Spodopterafrugiperda, fall armyworm; Diatraea grandiosella, southwestern cornborer; Elasmopalpus lignosellus, lesser cornstalk borer; Diatraeasaccharalis, surgarcane borer; western corn rootworm, e.g., Diabroticavirgifera virgifera; northern corn rootworm, e.g., Diabroticalongicornis barberi; southern corn rootworm, e.g., Diabroticaundecimpunctata howardi; Melanotus spp., wireworms; Cyclocephalaborealis, northern masked chafer (white grub); Cyclocephala immaculata,southern masked chafer (white grub); Popillia japonica, Japanese beetle;Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maizebillbug; Rhopalosiphum maidis, corn leaf aphid; Anuraphis maidiradicis,corn root aphid; Blissus leucopterus leucopterus, chinch bug; Melanoplusfemurrubrum, redlegged grasshopper; Melanoplus sanguinipes, migratorygrasshopper; Hylemya platura, seedcorn maggot; Agromyza parvicornis,corn blotch leafminer; Anaphothrips obscrurus, grass thrips; Solenopsismilesta, thief ant; Tetranychus urticae, two spotted spider mite;Sorghum: Chilo partellus, sorghum borer; Spodoptera frugiperda, fallarmyworm; Helicoverpa zea, corn earworm; Elasmopalpus lignosellus, lesercornstalk borer; Feltia subterranea, granulate cutworm; Phyllophagacrinita, white grub; Eleodes, Conoderus, and Aeolus spp., wireworms;Oulema melanopus, cereal leaf beetle; Chaetocnema pulicaria, corn fleabeetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis; cornleaf aphid; Sipha flava, yellow sugarcane aphid; chinch bug, e.g.,Blissus leucopterus leucopterus; Contarinia sorghicola, sorghum midge;Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae,two-spotted spider mite; Wheat: Pseudaletia unipunctata, army worm;Spodoptera frugiperda, fall armyworm; Elasmopalpus lignosellus, lessercornstalk borer; Agrotis orthogonia, pale western cutworm; Elasmopalpuslignosellus, lesser cornstalk borer; Oulema melanopus, cereal leafbeetle; Hypera punctata, clover leaf weevil; southern corn rootworm,e.g., Diabrotica undecimpunctata howardi; Russian wheat aphid;Schizaphis graminum, greenbug; Macrosiphum avenae, English grain aphid;Melanoplus femurrubrum, redlegged grasshopper; Melanoplusdifferentialis, differential grasshopper; Melanoplus sanguinipes,migratory grasshopper; Mayetiola destructor, Hessian fly; Sitodiplosismosellana, wheat midge; Meromyza americana, wheat stem maggot; Hylemyacoarctata, wheat bulb fly; Frankliniella fusca, tobacco thrips; Cephuscinctus, wheat stem sawfly; Aceria tulipae, wheat curl mite; Sunflower:Cylindrocupturus adspersus, sunflower stem weevil; Smicronyx fulus, redsunflower seed weevil; Smicronyx sordidus, gray sunflower seed weevil;Suleima helianthana, sunflower bud moth; Homoeosoma electellum,sunflower moth; Zygogramma exclamationis, sunflower beetle; Bothyrusgibbosus, carrot beetle; Neolasioptera murtfeldtiana, sunflower seedmidge; Cotton: Heliothis virescens, tobacco budworm; Helicoverpa zea,cotton bollworm; Spodoptera exigua, beet armyworm; Pectinophoragossypiella, pink bollworm; boll weevil, e.g., Anthonomus grandis; Aphisgossypii, cotton aphid; Pseudatomoscelis seriatus, cotton fleahopper;Trialeurodes abutilonea, bandedwinged whitefly; Lygus lineolaris,tarnished plant bug; Melanoplus femurrubrum, redlegged grasshopper;Melanoplus differentialis, differential grasshopper; Thrips tabaci,onion thrips; Franklinkiella fusca, tobacco thrips; Tetranychuscinnabarinus, carmine spider mite; Tetranychus urticae, two-spottedspider mite; Rice: Diatraea saccharalis, sugarcane borer; Spodopterafrugiperda, fall armyworm; Helicoverpa zea, corn earworm; Colaspisbrunnea, grape colaspis; Lissorhoptrus oryzophilus, rice water weevil;Sitophilus oryzae, rice weevil; Nephotettix nigropictus, rice leafhoper;chinch bug, e.g., Blissus leucopterus leucopterus; Acrosternum hilare,green stink bug; Soybean: Pseudoplusia includens, soybean looper;Anticarsia gemmatalis, velvetbean caterpillar; Plathypena scabra, greencloverworm; Ostrinia nubilalis, European corn borer; Agrotis ipsilon,black cutworm; Spodoptera exigua, beet armyworm; Heliothis virescens,tobacco budworm; Helicoverpa zea, cotton bollworm; Epilachna varivestis,Mexican bean beetle; Myzus persicae, green peach aphid; Empoasca fabae,potato leafhopper; Acrosternum hilare, green stink bug; Melanoplusfemurrubrum, redlegged grasshopper; Melanoplus differentialis,differential grasshopper; Hylemya platura, seedcorn maggot; Sericothripsvariabilis, soybean thrips; Thrips tabaci, onion thrips; Tetranychusturkestani, strawberry spider mite; Tetranychus urticae, two-spottedspider mite; Barley: Ostrinia nubilalis, European corn borer; Agrotisipsilon, black cutworm; Schizaphis graminum, greenbug; chinch bug, e.g.,Blissus leucopterus leucopterus; Acrosternum hilare, green stink bug;Euschistus servus, brown stink bug; Jylemya platura, seedcorn maggot;Mayetiola destructor, Hessian fly; Petrobia latens, brown wheat mite;Oil Seed Rape: Vrevicoryne brassicae, cabbage aphid; Phyllotretacruciferae, crucifer flea beetle; Phyllotreta striolata, striped fleabeetle; Phyllotreta nemorum, striped turnip flea beetle; Meligethesaeneus, rapeseed beetle; and the pollen beetles Meligethes rufimanus,Meligethes nigrescens, Meligethes canadianus, and Meligethesviridescens; Potato: Leptinotarsa decemlineata, Colorado potato beetle.

The methods and compositions provided herein may be effective againstHemiptera such as Lygus hesperus, Lygus lineolaris, Lygus pratensis,Lygus rugulipennis Popp, Lygus pabulinus, Calocoris norvegicus, Orthopscompestris, Plesiocoris rugicollis, Cyrtopeltis modestus, Cyrtopeltisnotatus, Spanagonicus albofasciatus, Diaphnocoris chlorinonis,Labopidicola allii, Pseudatomoscelis seriatus, Adelphocoris rapidus,Poecilocapsus lineatus, Blissus leucopterus, Nysius ericae, Nysiusraphanus, Euschistus servus, Nezara viridula, Eurygaster, Coreidae,Pyrrhocoridae, Tinidae, Blostomatidae, Reduviidae, and Cimicidae. Pestsof interest also include Araecerus fasciculatus, coffee bean weevil;Acanthoscelides obtectus, bean weevil; Bruchus rufmanus, broadbeanweevil; Bruchus pisorum, pea weevil; Zabrotes subfasciatus, Mexican beanweevil; Diabrotica balteata, banded cucumber beetle; Cerotomatrifurcata, bean leaf beetle; Diabrotica virgifera, Mexican cornrootworm; Epitrix cucumeris, potato flea beetle; Chaetocnema confinis,sweet potato flea beetle; Hypera postica, alfalfa weevil; Anthonomusquadrigibbus, apple curculio; Sternechus paludatus, bean stalk weevil;Hypera brunnipennis, Egyptian alfalfa weevil; Sitophilus granaries,granary weevil; Craponius inaequalis, grape curculio; Sitophiluszeamais, maize weevil; Conotrachelus nenuphar, plum curculio; Euscepespostfaciatus, West Indian sweet potato weevil; Maladera castanea,Asiatic garden beetle; Rhizotrogus majalis, European chafer;Macrodactylus subspinosus, rose chafer; Tribolium confusum, confusedflour beetle; Tenebrio obscurus, dark mealworm; Tribolium castaneum, redflour beetle; Tenebrio molitor, yellow mealworm.

Nematodes include parasitic nematodes such as root-knot, cyst, andlesion nematodes, including Heterodera spp., Meloidogyne spp., andGlobodera spp.; particularly members of the cyst nematodes, including,but not limited to, Heterodera glycines (soybean cyst nematode);Heterodera schachtii (beet cyst nematode); Heterodera avenae (cerealcyst nematode); and Globodera rostochiensis and Globodera pailida(potato cyst nematodes). Lesion nematodes include Pratylenchus spp.

Insect pests may be tested for pesticidal activity of compositions ofthe invention in early developmental stages, e.g., as larvae or otherimmature forms. The insects may be reared in total darkness at fromabout 20° C. to about 30° C. and from about 30% to about 70% relativehumidity. Bioassays may be performed as described in Czapla and Lang(1990) J. Econ. Entomol. 83 (6): 2480-2485. See, also the experimentalsection herein.

III. Expression Cassettes

Polynucleotides encoding the pesticidal proteins provided herein can beprovided in expression cassettes for expression in an organism ofinterest. The cassette will include 5′ and 3′ regulatory sequencesoperably linked to a polynucleotide encoding a pesticidal polypeptideprovided herein that allows for expression of the polynucleotide. Thecassette may additionally contain at least one additional gene orgenetic element to be cotransformed into the organism. Where additionalgenes or elements are included, the components are operably linked.Alternatively, the additional gene(s) or element(s) can be provided onmultiple expression cassettes. Such an expression cassette is providedwith a plurality of restriction sites and/or recombination sites forinsertion of the polynucleotides to be under the transcriptionalregulation of the regulatory regions. The expression cassette mayadditionally contain a selectable marker gene.

The expression cassette will include in the 5′-3′ direction oftranscription, a transcriptional and translational initiation region(i.e., a promoter), a pesticidal polynucleotide of the invention, and atranscriptional and translational termination region (i.e., terminationregion) functional in the organism of interest, i.e., a plant orbacteria. The promoters of the invention are capable of directing ordriving expression of a coding sequence in a host cell. The regulatoryregions (i.e., promoters, transcriptional regulatory regions, andtranslational termination regions) may be endogenous or heterologous tothe host cell or to each other. As used herein, “heterologous” inreference to a sequence is a sequence that originates from a foreignspecies, or, if from the same species, is substantially modified fromits native form in composition and/or genomic locus by deliberate humanintervention. As used herein, a chimeric gene comprises a codingsequence operably linked to a transcription initiation region that isheterologous to the coding sequence.

Convenient termination regions are available from the Ti-plasmid of A.tumefaciens, such as the octopine synthase and nopaline synthasetermination regions. See also Guerineau et al. (1991) Mol. Gen. Genet.262:141-144; Proudfoot (1991) Cell 64:671-674; Sanfacon et al. (1991)Genes Dev. 5:141-149; Mogen et al. (1990) Plant Cell 2:1261-1272; Munroeet al. (1990) Gene 91:151-158; Ballas et al. (1989) Nucleic Acids Res.17:7891-7903; and Joshi et al. (1987) Nucleic Acids Res. 15:9627-9639.

Additional regulatory signals include, but are not limited to,transcriptional initiation start sites, operators, activators,enhancers, other regulatory elements, ribosomal binding sites, aninitiation codon, termination signals, and the like. See, for example,U.S. Pat. Nos. 5,039,523 and 4,853,331; EPO 0480762A2; Sambrook et al.(1992) Molecular Cloning: A Laboratory Manual, ed. Maniatis et al. (ColdSpring Harbor Laboratory Press, Cold Spring Harbor, N.Y.), hereinafter“Sambrook 11”; Davis et al., eds. (1980) Advanced Bacterial Genetics(Cold Spring Harbor Laboratory Press), Cold Spring Harbor, N.Y., and thereferences cited therein.

In preparing the expression cassette, the various DNA fragments may bemanipulated, so as to provide for the DNA sequences in the properorientation and, as appropriate, in the proper reading frame. Towardthis end, adapters or linkers may be employed to join the DNA fragmentsor other manipulations may be involved to provide for convenientrestriction sites, removal of superfluous DNA, removal of restrictionsites, or the like. For this purpose, in vitro mutagenesis, primerrepair, restriction, annealing, resubstitutions, e.g., transitions andtransversions, may be involved.

A number of promoters can be used in the practice of the invention. Thepromoters can be selected based on the desired outcome. The nucleicacids can be combined with constitutive, inducible, tissue-preferred, orother promoters for expression in the organism of interest. See, forexample, promoters set forth in WO 99/43838 and in U.S. Pat. Nos:8,575,425; 7,790,846; 8,147,856; 8,586832; 7,772,369; 7,534,939;6,072,050; 5,659,026; 5,608,149; 5,608,144; 5,604,121; 5,569,597;5,466,785; 5,399,680; 5,268,463; 5,608,142; and 6,177,611; hereinincorporated by reference.

For expression in plants, constitutive promoters also include CaMV 35Spromoter (Odell et al. (1985) Nature 313:810-812); rice actin (McElroyet al. (1990) Plant Cell 2:163-171); ubiquitin (Christensen et al.(1989) Plant Mol. Biol. 12:619-632 and Christensen et al. (1992) PlantMol. Biol. 18:675-689); pEMU (Last et al. (1991) Theor. Appl. Genet.81:581-588); MAS (Velten et al. (1984) EMBO J. 3:2723-2730). Induciblepromoters include those that drive expression of pathogenesis-relatedproteins (PR proteins), which are induced following infection by apathogen. See, for example, Redolfi et al. (1983) Neth. J. Plant Pathol.89:245-254; Uknes et al. (1992) Plant Cell 4:645-656; and Van Loon(1985) Plant Mol. Virol. 4:111-116; and WO 99/43819, herein isincorporated by reference. Promoters that are expressed locally at ornear the site of pathogen infection may also be used (Marineau et al.(1987) Plant Mol. Biol. 9:335-342; Matton et al. (1989) MolecularPlant-Microbe Interactions 2:325-331; Somsisch et al. (1986) Proc. Natl.Acad. Sci. USA 83:2427-2430; Somsisch et al. (1988) Mol. Gen. Genet.2:93-98; and Yang (1996) Proc. Natl. Acad. Sci. USA 93:14972-14977; Chenet al. (1996) Plant J. 10:955-966; Zhang et al. (1994) Proc. Natl. Acad.Sci. USA 91:2507-2511; Warner et al. (1993) Plant J. 3:191-201; Siebertzet al. (1989) Plant Cell 1:961-968; Cordero et al. (1992) Physiol. Mol.Plant Path. 41:189-200; U.S. Pat. No. 5,750,386 (nematode-inducible);and the references cited therein).

Wound-inducible promoters may be used in the constructions of theinvention. Such wound-inducible promoters include pin II promoter (Ryan(1990) Ann. Rev. Phytopath. 28:425-449; Duan et al. (1996) NatureBiotechnology 14:494-498); wun1 and wun2 (U.S. Pat. No. 5,428,148); win1and win2 (Stanford et al. (1989) Mol. Gen. Genet. 215:200-208); systemin(McGurl et al. (1992) Science 225:1570-1573); WIP1 (Rohmeier et al.(1993) Plant Mol. Biol. 22:783-792; Eckelkamp et al. (1993) FEBS Letters323:73-76); MPI gene (Corderok et al. (1994) Plant J. 6(2):141-150); andthe like, herein incorporated by reference.

Tissue-preferred promoters for use in the invention include those setforth in Yamamoto et al. (1997) Plant J. 12(2):255-265; Kawamata et al.(1997) Plant Cell Physiol. 38(7):792-803; Hansen et al. (1997) Mol. GenGenet. 254(3):337-343; Russell et al. (1997) Transgenic Res.6(2):157-168; Rinehart et al. (1996) Plant Physiol. 112(3):1331-1341;Van Camp et al. (1996) Plant Physiol. 112(2):525-535; Canevascini et al.(1996) Plant Physiol. 112(2):513-524; Yamamoto et al. (1994) Plant CellPhysiol. 35(5):773-778; Lam (1994) Results Probl. Cell Differ.20:181-196; Orozco et al. (1993) Plant Mol Biol. 23(6):1129-1138;Matsuoka et al. (1993) Proc Natl. Acad. Sci. USA 90(20):9586-9590; andGuevara-Garcia et al. (1993) Plant J. 4(3):495-505.

Leaf-preferred promoters include those set forth in Yamamoto et al.(1997) Plant J. 12(2):255-265; Kwon et al. (1994) Plant Physiol.105:357-67; Yamamoto et al. (1994) Plant Cell Physiol. 35(5):773-778;Gotor et al. (1993) Plant 3:509-18; Orozco et al. (1993) Plant Mol.Biol. 23(6):1129-1138; and Matsuoka et al. (1993) Proc. Natl. Acad. Sci.USA 90(20):9586-9590.

Root-preferred promoters are known and include those in Hire et al.(1992) Plant Mol. Biol. 20(2):207-218 (soybean root-specific glutaminesynthetase gene); Keller and Baumgartner (1991) Plant Cell3(10):1051-1061 (root-specific control element); Sanger et al. (1990)Plant Mol. Biol. 14(3):433-443 (mannopine synthase (MAS) gene ofAgrobacterium tumefaciens); and Miao et al. (1991) Plant Cell 3(1):11-22(cytosolic glutamine synthetase (GS)); Bogusz et al. (1990) Plant Cell2(7):633-641; Leach and Aoyagi (1991) Plant Science (Limerick)79(1):69-76 (rolC and rolD); Teeri et al. (1989) EMBO J. 8(2):343-350;Kuster et al. (1995) Plant Mol. Biol. 29(4):759-772 (the VfENOD-GRP3gene promoter); and Capana et al. (1994) Plant Mol. Biol. 25(4):681-691(rolB promoter). See also U.S. Pat. Nos. 5,837,876; 5,750,386;5,633,363; 5,459,252; 5,401,836; 5,110,732; and 5,023,179.

“Seed-preferred” promoters include both “seed-specific” promoters (thosepromoters active during seed development such as promoters of seedstorage proteins) as well as “seed-germinating” promoters (thosepromoters active during seed germination). See Thompson et al. (1989)BioEssays 10:108. Seed-preferred promoters include, but are not limitedto, Cim1 (cytokinin-induced message); cZ19B1 (maize 19 kDa zein); milps(myo-inositol-1-phosphate synthase) (see Int'l Appl. Pub. No. WO00/11177 and U.S. Pat. No. 6,225,529). Gamma-zein is anendosperm-specific promoter. Globulin 1 (Glb-1) is a representativeembryo-specific promoter. For dicots, seed-specific promoters include,but are not limited to, bean β-phaseolin, napin, β-conglycinin, soybeanlectin, cruciferin, and the like. For monocots, seed-specific promotersinclude, but are not limited to, maize 15 kDa zein, 22 kDa zein, 27 kDazein, gamma-zein, waxy, shrunken 1, shrunken 2, Globulin 1, etc. Seealso Int'l Appl. Pub. No. WO 00/12733, where seed-preferred promotersfrom end1 and end2 genes are disclosed.

For expression in a bacterial host, promoters that function in bacteriaare well-known in the art. Such promoters include any of the knowncrystal protein gene promoters, including the promoters of any of thepesticidal proteins of the invention, and promoters specific for B.thuringiensis sigma factors. Alternatively, mutagenized or recombinantcrystal protein-encoding gene promoters may be recombinantly engineeredand used to promote expression of the novel gene segments disclosedherein.

The expression cassette can also comprise a selectable marker gene forthe selection of transformed cells. Selectable marker genes are utilizedfor the selection of transformed cells or tissues. Marker genes includegenes encoding antibiotic resistance, such as those encoding neomycinphosphotransferase II (NEO) and hygromycin phosphotransferase (HPT), aswell as genes conferring resistance to herbicidal compounds, such asglufosinate ammonium, bromoxynil, imidazolinones, and2,4-dichlorophenoxyacetate (2,4-D). Additional selectable markers areknown and any can be used in the practice of the invention. See, forexample, Int'l Appl. No. PCT/US2015/066648, filed on Dec. 18, 2015,herein incorporated by reference in its entirety, which disclosesglufosinate resistance sequences that can be employed as selectablemarkers.

IV. Methods, Host Cells, and Plant Cells

As indicated, DNA constructs comprising nucleotide sequences encodingthe pesticidal proteins or active variants or fragment thereof can beused to transform plants of interest or other organisms of interest.Methods for transformation involve introducing a nucleotide constructinto a plant. By “introducing” is intended to introduce the nucleotideconstruct to the plant or other host cell in such a manner that theconstruct gains access to the interior of a cell of the plant or hostcell. The methods of the invention do not require a particular methodfor introducing a nucleotide construct to a plant or host cell, onlythat the nucleotide construct gains access to the interior of at leastone cell of the plant or the host organism. Methods for introducingnucleotide constructs into plants and other host cells are known in theart including, but not limited to, stable transformation methods,transient transformation methods, and virus-mediated methods.

The methods result in a transformed organisms, such as a plant,including whole plants, as well as plant organs (e.g., leaves, stems,roots, etc.), seeds, plant cells, propagules, embryos and progeny of thesame. Plant cells can be differentiated or undifferentiated (e.g.callus, suspension culture cells, protoplasts, leaf cells, root cells,phloem cells, and pollen).

“Transgenic plants” or “transformed plants” or “stably transformed”plants or cells or tissues refers to plants that have incorporated orintegrated a polynucleotide encoding at least one pesticidal polypeptideof the invention. It is recognized that other exogenous or endogenousnucleic acid sequences or DNA fragments may also be incorporated intothe plant cell. Agrobacterium- and biolistic-mediated transformationremain the two predominantly employed approaches. However,transformation may be performed by infection, transfection,microinjection, electroporation, microprojection, biolistics or particlebombardment, electroporation, silica/carbon fibers, ultrasound mediated,PEG mediated, calcium phosphate co-precipitation, polycation DMSOtechnique, DEAE dextran procedure, Agro and viral mediated(Caulimoriviruses, Geminiviruses, RNA plant viruses), liposome mediated,and the like.

Transformation protocols as well as protocols for introducingpolypeptides or polynucleotide sequences into plants may vary dependingon the type of plant or plant cell, i.e., monocot or dicot, targeted fortransformation. Methods for transformation are known in the art andinclude those set forth in U.S. Pat. Nos: 8,575,425; 7,692,068;8,802,934; and 7,541,517; each of which is herein incorporated byreference. See, also, Rakoczy-Trojanowska, M. (2002) Cell Mol Biol Lett.7:849-858; Jones et al. (2005) Plant Methods 1:5; Rivera et al. (2012)Physics of Life Reviews 9:308-345; Bartlett et al. (2008) Plant Methods4:1-12; Bates, G. W. (1999) Methods in Molecular Biology 111:359-366;Binns and Thomashow (1988) Annual Reviews in Microbiology 42:575-606;Christou, P. (1992) The Plant Journal 2:275-281; Christou, P. (1995)Euphytica 85:13-27; Tzfira et al. (2004) TRENDS in Genetics 20:375-383;Yao et al. (2006) Journal of Experimental Botany 57:3737-3746; Zupan andZambryski (1995) Plant Physiology 107:1041-1047; and Jones et al. (2005)Plant Methods 1:5.

Transformation may result in stable or transient incorporation of thenucleic acid into the cell. “Stable transformation” is intended to meanthat the nucleotide construct introduced into a host cell integratesinto the genome of the host cell and is capable of being inherited bythe progeny thereof. “Transient transformation” is intended to mean thata polynucleotide is introduced into the host cell and does not integrateinto the genome of the host cell.

Methods for transformation of chloroplasts are known in the art. See,for example, Svab et al. (1990) Proc. Natl. Acad. Sci. USA 87:8526-8530;Svab and Maliga (1993) Proc. Natl. Acad. Sci. USA 90:913-917; Svab andMaliga (1993) EMBO J. 12:601-606. The method relies on particle gundelivery of DNA containing a selectable marker and targeting of the DNAto the plastid genome through homologous recombination. Additionally,plastid transformation can be accomplished by transactivation of asilent plastid-borne transgene by tissue-preferred expression of anuclear-encoded and plastid-directed RNA polymerase. Such a system hasbeen reported in McBride et al. (1994) Proc. Natl. Acad. Sci. USA91:7301-7305.

The cells that have been transformed may be grown into plants inaccordance with conventional ways. See, for example, McCormick et al.(1986) Plant Cell Reports 5:81-84. These plants may then be grown, andeither pollinated with the same transformed strain or different strains,and the resulting hybrid having constitutive expression of the desiredphenotypic characteristic identified. Two or more generations may begrown to ensure that expression of the desired phenotypic characteristicis stably maintained and inherited and then seeds harvested to ensureexpression of the desired phenotypic characteristic has been achieved.In this manner, the present invention provides transformed seed (alsoreferred to as “transgenic seed”) having a nucleotide construct of theinvention, for example, an expression cassette of the invention, stablyincorporated into their genome.

In specific embodiments, the sequences provide herein can be targeted tospecific sites within the genome of the host cell or plant cell. Suchmethods include, but are not limited to, meganucleases designed againstthe plant genomic sequence of interest (D'Halluin et al. 2013 PlantBiotechnol J); CRISPR-Cas9, TALENs, and other technologies for preciseediting of genomes (Feng, et al. Cell Research 23:1229-1232, 2013,Podevin, et al. Trends Biotechnology, online publication, 2013, Wei etal., J Gen Genomics, 2013, Zhang et al (2013) WO 2013/026740); Cre-loxsite-specific recombination (Dale et al. (1995) Plant J 7:649-659;Lyznik, et al. (2007) Transgenic Plant J 1:1-9; FLP-FRT recombination(Li et al. (2009) Plant Physiol 151:1087-1095); Bxb1-mediatedintegration (Yau et al. Plant J (2011) 701:147-166); zinc-fingermediated integration (Wright et al. (2005) Plant J 44:693-705); Cai etal. (2009) Plant Mol Biol 69:699-709); and homologous recombination(Lieberman-Lazarovich and Levy (2011) Methods Mol Biol 701: 51-65);Puchta (2002) Plant Mol Biol 48:173-182).

The sequence provided herein may be used for transformation of any plantspecies, including, but not limited to, monocots and dicots. Examples ofplants of interest include, but are not limited to, corn (maize),sorghum, wheat, sunflower, tomato, crucifers, peppers, potato, cotton,rice, soybean, sugarbeet, sugarcane, tobacco, barley, and oilseed rape,Brassica sp., alfalfa, rye, millet, safflower, peanuts, sweet potato,cassaya, coffee, coconut, pineapple, citrus trees, cocoa, tea, banana,avocado, fig, guava, mango, olive, papaya, cashew, macadamia, almond,oats, vegetables, ornamentals, and conifers.

Vegetables include, but are not limited to, tomatoes, lettuce, greenbeans, lima beans, peas, and members of the genus Curcumis such ascucumber, cantaloupe, and musk melon. Ornamentals include, but are notlimited to, azalea, hydrangea, hibiscus, roses, tulips, daffodils,petunias, carnation, poinsettia, and chrysanthemum. Preferably, plantsof the present invention are crop plants (for example, maize, sorghum,wheat, sunflower, tomato, crucifers, peppers, potato, cotton, rice,soybean, sugarbeet, sugarcane, tobacco, barley, oilseed rape, etc.).

As used herein, the term plant includes plant cells, plant protoplasts,plant cell tissue cultures from which plants can be regenerated, plantcalli, plant clumps, and plant cells that are intact in plants or partsof plants such as embryos, pollen, ovules, seeds, leaves, flowers,branches, fruit, kernels, ears, cobs, husks, stalks, roots, root tips,anthers, and the like. Grain is intended to mean the mature seedproduced by commercial growers for purposes other than growing orreproducing the species. Progeny, variants, and mutants of theregenerated plants are also included within the scope of the invention,provided that these parts comprise the introduced polynucleotides.Further provided is a processed plant product or byproduct that retainsthe sequences disclosed herein, including for example, soymeal.

In another embodiment, the genes encoding the pesticidal proteins can beused to transform insect pathogenic organisms. Such organisms includebaculoviruses, fungi, is protozoa, bacteria, and nematodes.Microorganism hosts that are known to occupy the “phytosphere”(phylloplane, phyllosphere, rhizosphere, and/or rhizoplana) of one ormore crops of interest may be selected. These microorganisms areselected so as to be capable of successfully competing in the particularenvironment with the wild-type microorganisms, provide for stablemaintenance and expression of the gene expressing the pesticidalprotein, and desirably, provide for improved protection of the pesticidefrom environmental degradation and inactivation.

Such microorganisms include archaea, bacteria, algae, and fungi. Ofparticular interest are microorganisms such as bacteria, e.g., Bacillus,Pseudomonas, Erwinia, Serratia, Klebsiella, Xanthomonas, Streptomyces,Rhizobium, Rhodopseudomonas, Methylius, Agrobacterium, Acetobacter,Lactobacillus, Arthrobacter, Azotobacter, Leuconostoc, and Alcaligenes.Fungi include yeast, e.g., Saccharomyces, Cryptococcus, Kluyveromyces,Sporobolomyces, Rhodotorula, and Aureobasidium. Of particular interestare such phytosphere bacterial species as Pseudomonas syringae,Pseudomonas aeruginosa, Pseudomonas fluorescens, Serratia marcescens,Acetobacter xylinum, Agrobacteria, Rhodopseudomonas spheroides,Xanthomonas campestris, Rhizobium melioti, Alcaligenes entrophus,Clavibacter xyli and Azotobacter vinlandir and phytosphere yeast speciessuch as Rhodotorula rubra, R. glutinis, R. marina, R. aurantiaca,Cryptococcus albidus, C. diffluens, C. laurentii, Saccharomyces rosei,S. pretoriensis, S. cerevisiae, Sporobolomyces rosues, S. odorus,Kluyveromyces veronae, Aureobasidium pollulans, Bacillus thuringiensis,Escherichia coli, Bacillus subtilis, and the like.

Illustrative prokaryotes, both Gram-negative and gram-positive, includeEnterobacteriaceae, such as Escherichia, Erwinia, Shigella, Salmonella,and Proteus; Bacillaceae; Rhizobiceae, such as Rhizobium; Spirillaceae,such as photobacterium, Zymomonas, Serratia, Aeromonas, Vibrio,Desulfovibrio, Spirillum; Lactobacillaceae; Pseudomonadaceae, such asPseudomonas and Acetobacter; Azotobacteraceae and Nitrobacteraceae.Fungi include Phycomycetes and Ascomycetes, e.g., yeast, such asSaccharomyces and Schizosaccharomyces; and Basidiomycetes yeast, such asRhodotorula, Aureobasidium, Sporobolomyces, and the like.

Genes encoding pesticidal proteins can be introduced by means ofelectrotransformation, PEG induced transformation, heat shock,transduction, conjugation, and the like. Specifically, genes encodingthe pesticidal proteins can be cloned into a shuttle vector, forexample, pHT3101 (Lerecius et al. (1989) FEMS Microbiol. Letts. 60:211-218. The shuttle vector pHT3101 containing the coding sequence forthe particular pesticidal protein gene can, for example, be transformedinto the root-colonizing Bacillus by means of electroporation (Lereciuset al. (1989) FEMS Microbiol. Letts. 60: 211-218).

Expression systems can be designed so that pesticidal proteins aresecreted outside the cytoplasm of gram-negative bacteria by fusing anappropriate signal peptide to the amino-terminal end of the pesticidalprotein. Signal peptides recognized by E. coli include the OmpA protein(Ghrayeb et al. (1984) EMBO J, 3: 2437-2442).

Pesticidal proteins and active variants thereof can be fermented in abacterial host and the resulting bacteria processed and used as amicrobial spray in the same manner that Bacillus thuringiensis strainshave been used as insecticidal sprays. In the case of a pesticidalprotein(s) that is secreted from Bacillus, the secretion signal isremoved or mutated using procedures known in the art. Such mutationsand/or deletions prevent secretion of the pesticidal protein(s) into thegrowth medium during the fermentation process. The pesticidal proteinsare retained within the cell, and the cells are then processed to yieldthe encapsulated pesticidal proteins.

Alternatively, the pesticidal proteins are produced by introducingheterologous genes into a cellular host. Expression of the heterologousgene results, directly or indirectly, in the intracellular productionand maintenance of the pesticide. These cells are then treated underconditions that prolong the activity of the toxin produced in the cellwhen the cell is applied to the environment of target pest(s). Theresulting product retains the toxicity of the toxin. These naturallyencapsulated pesticidal proteins may then be formulated in accordancewith conventional techniques for application to the environment hostinga target pest, e.g., soil, water, and foliage of plants. See, forexample, U.S. Pat. No. 6,468,523 and U.S. Publication No. 20050138685,and the references cited therein. In the present invention, atransformed microorganism (which is includes whole organisms, cells,spore(s), pesticidal protein(s), pesticidal component(s), pest-impactingcomponent(s), mutant(s), living or dead cells and cell components,including mixtures of living and dead cells and cell components, andincluding broken cells and cell components) or an isolated pesticidalprotein can be formulated with an acceptable carrier into a pesticidalor agricultural composition(s) that is, for example, a suspension, asolution, an emulsion, a dusting powder, a dispersible granule, awettable powder, and an emulsifiable concentrate, an aerosol, animpregnated granule, an adjuvant, a coatable paste, and alsoencapsulations in, for example, polymer substances.

Agricultural compositions may comprise a polypeptide, a recombinogenicpolypeptide or a variant or fragment thereof, as disclosed herein. Theagricultural composition disclosed herein may be applied to theenvironment of a plant or an area of cultivation, or applied to theplant, plant part, plant cell, or seed.

Such compositions disclosed above may be obtained by the addition of asurface-active agent, an inert carrier, a preservative, a humectant, afeeding stimulant, an attractant, an encapsulating agent, a binder, anemulsifier, a dye, a UV protectant, a buffer, a flow agent orfertilizers, micronutrient donors, or other preparations that influenceplant growth. One or more agrochemicals including, but not limited to,herbicides, insecticides, fungicides, bactericides, nematicides,molluscicides, acaracides, plant growth regulators, harvest aids, andfertilizers, can be combined with carriers, surfactants or adjuvantscustomarily employed in the art of formulation or other components tofacilitate product handling and application for particular target pests.Suitable carriers and adjuvants can be solid or liquid and correspond tothe substances ordinarily employed in formulation technology, e.g.,natural or regenerated mineral substances, solvents, dispersants,wetting agents, tackifiers, binders, or fertilizers. The activeingredients of the present invention are normally applied in the form ofcompositions and can be applied to the crop area, plant, or seed to betreated. For example, the compositions of the present invention may beapplied to grain in preparation for or during storage in a grain bin orsilo, etc. The compositions of the present invention may be appliedsimultaneously or in succession with other compounds. Methods ofapplying an active ingredient of the present invention or anagrochemical composition of the present invention that contains at leastone of the pesticidal proteins produced by the bacterial strains of thepresent invention include, but are not limited to, foliar application,seed coating, and soil application. The number of applications and therate of application depend on the intensity of infestation by thecorresponding pest.

Suitable surface-active agents include, but are not limited to, anioniccompounds such as a carboxylate of, for example, a metal; a carboxylateof a long chain fatty acid; an N-acylsarcosinate; mono or di-esters ofphosphoric acid with fatty alcohol ethoxylates or salts of such esters;fatty alcohol sulfates such as sodium dodecyl sulfate, sodium octadecylsulfate or sodium cetyl sulfate; ethoxylated fatty alcohol sulfates;ethoxylated alkylphenol sulfates; lignin sulfonates; petroleumsulfonates; alkyl aryl sulfonates such as alkyl-benzene sulfonates orlower alkylnaphtalene sulfonates, e.g., butyl-naphthalene sulfonate;salts of sulfonated naphthalene-formaldehyde condensates; salts ofsulfonated phenol-formaldehyde condensates; more complex sulfonates suchas the amide sulfonates, e.g., the sulfonated condensation product ofoleic acid and N-methyl taurine; or the dialkyl sulfosuccinates, e.g.,the sodium sulfonate of dioctyl succinate. Non-ionic agents includecondensation products of fatty acid esters, fatty alcohols, fatty acidamides or fatty-alkyl- or alkenyl-substituted phenols with ethyleneoxide, fatty esters of polyhydric alcohol ethers, e.g., sorbitan fattyacid esters, condensation products of such esters with ethylene oxide,e.g., polyoxyethylene sorbitar fatty acid esters, block copolymers ofethylene oxide and propylene oxide, acetylenic glycols such as2,4,7,9-tetraethyl-5-decyn-4,7-diol, or ethoxylated acetylenic glycols.Examples of a cationic surface-active agent include, for instance, analiphatic mono-, di-, or polyamine such as an acetate, naphthenate oroleate; or oxygen-containing amine such as an amine oxide ofpolyoxyethylene alkylamine; an amide-linked amine prepared by thecondensation of a carboxylic acid with a di- or polyamine; or aquaternary ammonium salt.

Examples of inert materials include but are not limited to inorganicminerals such as kaolin, phyllosilicates, carbonates, sulfates,phosphates, or botanical materials such as cork, powdered corncobs,peanut hulls, rice hulls, and walnut shells.

The compositions of the present invention can be in a suitable form fordirect application or as a concentrate of primary composition thatrequires dilution with a suitable quantity of water or other diluantbefore application. The pesticidal concentration will vary dependingupon the nature of the particular formulation, specifically, whether itis a concentrate or to be used directly. The composition contains 1 to98% of a solid or liquid inert carrier, and 0 to 50% or 0.1 to 50% of asurfactant. These compositions will be administered at the labeled ratefor the commercial product, for example, about 0.01 lb-5.0 lb. per acrewhen in dry form and at about 0.01 pts.-10 pts. per acre when in liquidform.

In a further embodiment, the compositions, as well as the transformedmicroorganisms and pesticidal proteins, provided herein can be treatedprior to formulation to prolong the pesticidal activity when applied tothe environment of a target pest as long as the pretreatment is notdeleterious to the pesticidal activity. Such treatment can be bychemical and/or physical means as long as the treatment does notdeleteriously affect the properties of the composition(s). Examples ofchemical reagents include but are not limited to halogenating agents;aldehydes such as formaldehyde and glutaraldehyde; anti-infectives, suchas zephiran chloride; alcohols, such as isopropanol and ethanol; andhistological fixatives, such as Bouin's fixative and Helly's fixative(see, for example, Humason (1967) Animal Tissue Techniques (W.H. Freemanand Co.).

In one aspect, pests may be killed or reduced in numbers in a given areaby application of the pesticidal proteins of the invention to the area.Alternatively, the pesticidal proteins may be prophylactically appliedto an environmental area to prevent infestation by a susceptible pest.Preferably the pest ingests, or is contacted with, apesticidally-effective amount of the polypeptide. By“pesticidally-effective amount” is intended an amount of the pesticidethat is able to bring about death to at least one pest, or to noticeablyreduce pest growth, feeding, or normal physiological development. Thisamount will vary depending on such factors as, for example, the specifictarget pests to be controlled, the specific environment, location,plant, crop, or agricultural site to be treated, the environmentalconditions, and the method, rate, concentration, stability, and quantityof application of the pesticidally-effective polypeptide composition.The formulations or compositions may also vary with respect to climaticconditions, environmental considerations, and/or frequency ofapplication and/or severity of pest infestation.

The active ingredients are normally applied in the form of compositionsand can be applied to the crop area, plant, or seed to be treated.Methods are therefore provided for providing to a plant, plant cell,seed, plant part or an area of cultivation, an effective amount of theagricultural composition comprising the polypeptide, recombinogenicpolypeptide or an active variant or fragment thereof. By “effectiveamount” is intended an amount of a protein or composition sufficient tokill or control the pest or result in a noticeable reduction in pestgrowth, feeding, or normal physiological development. Such decreases inpest numbers, pest growth, pest feeding or pest normal development cancomprise any statistically significant decrease, including, for examplea decrease of about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%,55%, 60%, 65%, 70%, 75%, 85%, 90%, 95% or greater. For example, thecompositions may be applied to grain in preparation for or duringstorage in a grain bin or silo, etc. The compositions may be appliedsimultaneously or in succession with other compounds. Methods ofapplying an active ingredient or an agrochemical composition comprisingat least one of the polypeptides, recombinogenic polypeptides orvariants or fragments thereof as disclosed herein, include but are notlimited to, foliar application, seed coating, and soil application.

Methods for increasing plant yield are provided. The methods compriseproviding a plant or plant cell expressing a polynucleotide encoding thepesticidal polypeptide sequence disclosed herein and growing the plantor a seed thereof in a field infested with (or susceptible toinfestation by) a pest against which said polypeptide has pesticidalactivity. In some embodiments, the polypeptide has pesticidal activityagainst a lepidopteran, coleopteran, dipteran, hemipteran, or nematodepest, and said field is infested with a lepidopteran, hemipteran,coleopteran, dipteran, or nematode pest. As defined herein, the “yield”of the plant refers to the quality and/or quantity of biomass producedby the plant. By “biomass” is intended any measured plant product. Anincrease in biomass production is any improvement in the yield of themeasured plant product. Increasing plant yield has several commercialapplications. For example, increasing plant leaf biomass may increasethe yield of leafy vegetables for human or animal consumption.Additionally, increasing leaf biomass can be used to increase productionof plant-derived pharmaceutical or industrial products. An increase inyield can comprise any statistically significant increase including, butnot limited to, at least a 1% increase, at least a 3% increase, at leasta 5% increase, at least a 10% increase, at least a 20% increase, atleast a 30%, at least a 50%, at least a 70%, at least a 100% or agreater increase in yield compared to a plant not expressing thepesticidal sequence. In specific methods, plant yield is increased as aresult of improved pest resistance of a plant expressing a pesticidalprotein disclosed herein. Expression of the pesticidal protein resultsin a reduced ability of a pest to infest or feed.

The plants can also be treated with one or more chemical compositions,including one or more herbicide, insecticides, or fungicides.

Non-limiting embodiments include:

1. An isolated polypeptide having insecticidal activity, comprising

(a) a polypeptide comprising an amino acid sequence selected from thegroup consisting of sequences set forth in SEQ ID NOs: 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61,62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79,80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112,113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126,127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140,141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154,155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168,169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196,197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210,211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224,225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238,239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252,253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266,267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280,281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294,295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308,309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322,323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336,337, 338, 339, 340, and/or 341; or

(b) a polypeptide comprising an amino acid sequence having at least thepercent sequence identity set forth in Table 1 to an amino acid sequenceselected from the group consisting of sequences set forth in SEQ ID NOs:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38,39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56,57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92,93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108,109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122,123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136,137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150,151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164,165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178,179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192,193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206,207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220,221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234,235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248,249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262,263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276,277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290,291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304,305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318,319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332,333, 334, 335, 336, 337, 338, 339, 340, and/or 341.

2. The polypeptide of embodiment 1, wherein said polypeptide comprisesthe amino acid sequence set forth in SEQ ID Nos. 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44,45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62,63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98,99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112,113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126,127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140,141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154,155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168,169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196,197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210,211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224,225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238,239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252,253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266,267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280,281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294,295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308,309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322,323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336,337, 338, 339, 340, and/or 341.

3. The polypeptide of embodiment 1 or 2, further comprising heterologousamino acid sequences.

4. A composition comprising the polypeptide of any of embodiments 1, 2,or 3.

5. A recombinant nucleic acid molecule that encodes the polypeptide anyone of embodiments 1-3, wherein said recombinant nucleic acid moleculeis not the naturally occurring sequence encoding said polypeptide.

6. The recombinant nucleic acid of embodiment 5, wherein said nucleicacid molecule is a synthetic sequence that has been designed forexpression in a plant.

7. The recombinant nucleic acid molecule of embodiment 5 or 6, whereinsaid nucleic acid molecule is operably linked to a promoter capable ofdirecting expression in a plant cell.

8. The recombinant nucleic acid molecule of embodiment 5 or 6, whereinsaid nucleic acid molecule is operably linked to a promoter capable ofdirecting expression in a bacteria.

9. A host cell that contains the recombinant nucleic acid molecule ofany one of embodiments 5-8.

10. The host cell of embodiment 9, wherein said host cell is a bacterialhost cell.

11. A DNA construct comprising a promoter that drives expression in aplant cell operably linked to a recombinant nucleic acid moleculecomprising

(a) a nucleotide sequence that encodes a polypeptide comprising theamino acid sequence of any one of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45,46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63,64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99,100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113,114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127,128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141,142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155,156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169,170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183,184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197,198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211,212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225,226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239,240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253,254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267,268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281,282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295,296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309,310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323,324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337,338, 339, 340, and/or 341; or,

(b) a nucleotide sequence that encodes a polypeptide comprising an aminoacid sequence having at least the percent sequence identity set forth inTable 1 to an amino acid sequence selected from the group consisting ofsequences set forth in SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47,48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65,66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100,101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114,115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128,129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142,143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156,157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170,171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184,185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198,199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226,227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240,241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254,255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268,269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282,283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296,297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310,311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324,325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338,339, 340, and/or 341.

12. The DNA construct of embodiment 11, wherein said nucleotide sequenceis a synthetic DNA sequence that has been designed for expression in aplant.

13. A vector comprising the DNA construct of embodiment 11.

14. A host cell that contains the DNA construct of embodiment 11 or 12or the vector of claim 13.

15. The host cell of embodiment 14, wherein the host cell is a plantcell.

16. A transgenic plant comprising the host cell of embodiment 14 or 15.

17. A composition comprising the host cell of any one of embodiments 9,10, 14, or 15.

18. The composition of embodiment 17, wherein said composition isselected from the group consisting of a powder, dust, pellet, granule,spray, emulsion, colloid, and solution.

19. The composition of embodiment 17 or 18, wherein said compositioncomprises from about 1% to about 99% by weight of said polypeptide.

20. A method for controlling a pest population comprising contactingsaid population with a pesticidal-effective amount of the composition ofany one of embodiments 17-19.

21. A method for killing a pest population comprising contacting saidpopulation with a pesticidal-effective amount of the composition of anyone of embodiments 17-19.

22. A method for producing a polypeptide with pesticidal activity,comprising culturing the host cell of any one of embodiments 9, 10, 14,or 15 under conditions in which the nucleic acid molecule encoding thepolypeptide is expressed.

23. A plant having stably incorporated into its genome a DNA constructcomprising a nucleotide sequence that encodes a protein havingpesticidal activity, wherein said nucleotide sequence comprise

(a) a nucleotide sequence that encodes a polypeptide comprising theamino acid sequence of any one of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45,46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63,64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99,100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113,114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127,128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141,142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155,156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169,170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183,184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197,198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211,212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225,226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239,240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253,254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267,268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281,282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295,296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309,310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323,324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337,338, 339, 340, and/or 341; or,

(b) a nucleotide sequence that encodes a polypeptide comprising an aminoacid sequence having at least the percent sequence identity set forth inTable 1 to an amino acid sequence selected from the group consisting ofsequences set forth in SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47,48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65,66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100,101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114,115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128,129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142,143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156,157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170,171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184,185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198,199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226,227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240,241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254,255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268,269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282,283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296,297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310,311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324,325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338,339, 340, and/or 341.

24. A transgenic seed of the plant of embodiment 23.

25. A method for protecting a plant from an insect pest, comprisingexpressing in a plant or cell thereof a nucleotide sequence that encodesa pesticidal polypeptide, wherein said nucleotide sequence comprising

(a) a nucleotide sequence that encodes a polypeptide comprising theamino acid sequence of any one of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45,46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63,64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99,100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113,114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127,128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141,142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155,156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169,170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183,184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197,198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211,212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225,226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239,240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253,254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267,268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281,282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295,296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309,310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323,324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337,338, 339, 340, and/or 341; or,

(b) a nucleotide sequence that encodes a polypeptide comprising an aminoacid sequence having at least the percent sequence identity set forth inTable 1 to an amino acid sequence selected from the group consisting ofsequences set forth in SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47,48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65,66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100,101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114,115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128,129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142,143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156,157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170,171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184,185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198,199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226,227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240,241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254,255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268,269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282,283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296,297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310,311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324,325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338,339, 340, and/or 341.

26. The method of embodiment 25, wherein said plant produces apesticidal polypeptide having pesticidal against a lepidopteran pest, acoleopteran pest, or a hemipteran pest.

27. A method for increasing yield in a plant comprising growing in afield a plant or seed thereof having stably incorporated into its genomea DNA construct comprising a promoter that drives expression in a plantoperably linked to a nucleotide sequence that encodes a pesticidalpolypeptide, wherein said nucleotide sequence comprises

(a) a nucleotide sequence that encodes a polypeptide comprising theamino acid sequence of any one of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45,46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63,64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99,100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113,114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127,128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141,142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155,156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169,170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183,184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197,198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211,212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225,226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239,240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253,254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267,268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281,282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295,296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309,310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323,324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337,338, 339, 340, and/or 341; or

(b) a nucleotide sequence that encodes a polypeptide comprising an aminoacid sequence having at least the percent sequence identity set forth inTable 1 to an amino acid sequence selected from the group consisting ofsequences set forth in SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47,48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65,66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100,101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114,115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128,129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142,143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156,157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170,171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184,185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198,199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226,227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240,241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254,255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268,269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282,283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296,297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310,311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324,325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338,339, 340, and/or 341.

28. A method of obtaining a polynucleotide that encodes an improvedpolypeptide comprising pesticidal activity is provided, wherein theimproved polypeptide has at least one improved property over any one ofSEQ ID NOS: 1-341 comprising:

(a) recombining a plurality of parental polynucleotides comprising SEQID NO: 1-341 or an active variant or fragment thereof to produce alibrary of recombinant polynucleotides encoding recombinant pesticidalpolypeptides;

(b) screening the library to identify a recombinant polynucleotide thatencodes an improved recombinant pesticidal polypeptide that has anenhanced property improved over the parental polynucleotide;

(c) recovering the recombinant polynucleotide that encodes the improvedrecombinant pesticidal polypeptide identified in (b); and,

(d) repeating steps (a), (b) and (c) using the recombinantpolynucleotide recovered in step (c) as one of the plurality of parentalpolynucleotides in repeated step (a).

The following examples are offered by way of illustration and not by wayof limitation.

EXPERIMENTAL Example 1 Discovery of Novel Genes by Sequencing and DNAAnalysis

Microbial cultures were grown in liquid culture in standard laboratorymedia. Cultures were grown to saturation (16 to 24 hours) before DNApreparation. DNA was extracted from bacterial cells by detergent lysis,followed by binding to a silica matrix and washing with an ethanolbuffer. Purified DNA was eluted from the silica matrix with a mildlyalkaline aqueous buffer.

DNA for sequencing was tested for purity and concentration byspectrophotometry. Sequencing libraries were prepared using the NexteraXT library preparation kit according to the manufacturer's protocol.Sequence data was generated on a HiSeq 2000 according to the IlluminaHiSeq 2000 System User Guide protocol.

Sequencing reads were assembled into draft genomes using the CLC BioAssembly Cell software package. Following assembly, gene calls were madeby several methods and resulting gene sequences were interrogated toidentify novel homologs of pesticidal genes. Novel genes were identifiedby BLAST, by domain composition, and by pairwise alignment versus atarget set of pesticidal genes. A summary of such sequences is set forthin Table 1.

Genes identified in the homology search were amplified from bacterialDNA by PCR and cloned into bacterial expression vectors containing fusedin-frame purification tags. Cloned genes were expressed in E. coli andpurified by column chromatography. Purified proteins were assessed ininsect diet bioassay studies to identify active proteins.

Insect diet bioassays were performed using a wheat germ and agarartificial diet to which purified protein were applied as a surfacetreatment. Insect larvae were applied to treated diet and monitored formortality.

Insect diet bioassays were performed using a sucrose liquid dietcontained in a membrane sachet to which purified protein was added.Insect nymphs were allowed to feed on the diet sachet and were monitoredfor mortality. Insects tested in bioassays included the Brown Stink Bug(BSB), Euschistus servus, and the Southern Green Stink Bug (SGSB),Nezara viridula.

Example 2 Heterologous Expression in E. Coli

Each open reading frame set forth in Tables 3 and 4 was cloned into anE. coli expression vector containing a maltose binding protein (pMBP).The expression vector was transformed into BL21*RIPL. An LB culturesupplemented with carbenicillin was inoculated with a single colony andgrown overnight at 37° C. using 0.5% of the overnight culture, a freshculture was inoculated and grown to logarithmic phase at 37° C. Theculture was induced using 250 mM IPTG for 18 hours at 16° C. The cellswere pelleted and resuspended in 10 mM Tris pH7.4 and 150 mM NaClsupplemented with protease inhibitors. The protein expression wasevaluated by SDS-PAGE.

Example 3 Pesticidal Activity Against Coleopteran and Lepidoptera

Protein Expression: Each sequence set forth in Table 3 was expressed inE. coli as described in Example 2. 400 mL of LB was inoculated and grownto an OD600 of 0.6. The culture was induced with 0.25 mM IPTG overnightat 16° C. The cells were spun down and the cell pellet was resuspend in5 mL of buffer. The resuspension was sonicated for 2 min on ice.

Bioassay: Fall army worm (FAW), corn ear worm (CEW), European corn borer(ECB) southwestern corn borer (SWCB) and diamond backed moth (DBM or Px)eggs were purchased from a commercial insectary (Benzon Research Inc.,Carlisle, Pa.). The is FAW, CEW, ECB and BCW eggs were incubated to thepoint that eclosion would occur within 12 hrs of the assay setup. SWCBand DBM were introduced to the assay as neonate larvae. Assays werecarried out in 24-well trays containing multispecies lepidopteran diet(Southland Products Inc., Lake Village, Ark.). Samples of the sonicatedlysate were applied to the surface of the diet (diet overlay) andallowed to evaporate and soak into the diet. For CEW, FAW, BCW, ECB andSWCB, a 125 μl of sonicated lysate was added to the diet surface anddried. For DBM, 50 μl of a 1:2 dilution of sonicated lysate was added tothe diet surface. The bioassay plates were sealed with a plate sealingfilm vented with pin holes. The plates were incubated at 26° C. at 65%relative humidity (RH) on a 16:8 day:night cycle in a Percival for 5days. The assays were assessed for level of mortality, growth inhibitionand feeding inhibition.

For the western corn rootworm bioassay, the protein construct/lysate wasevaluated in an insect bioassay by dispensing 60 μl volume on the topsurface of diet in well/s of 24-well plate (Cellstar, 24-well, GreinerBio One) and allowed to dry. Each well contained 500 μl diet (Marrone etal., 1985). Fifteen to twenty neonate larvae were introduced in eachwell using a fine tip paint brush and the plate was covered withmembrane (Viewseal, Greiner Bio One). The bioassay was stored at ambienttemperature and scored for mortality, and/or growth/feeding inhibitionat day 4.

For Colorado Potato Beetle (CPB) a cork bore size No. 8 leaf disk wasexcised from potato leaf and was dipped in the protein construct/lysateuntil thoroughly wet and placed on top of filter disk (Millipore, glassfiber filter, 13 mm). 60 μl dH2O was added to each filter disk andplaced in each well of 24-well plate (Cellstar, 24-well, Greiner BioOne). The leaf disk was allowed to dry and five to seven first instarlarvae were introduced in each well using a fine tip paint brush. Theplate was covered with membrane (Viewseal, Greiner Bio One) and smallhole was punctured in each well of the membrane. The construct wasevaluated with four replicates, and scored for mortality and leaf damageon day 3.

Table 3 provides a summary of pesticidal activity against coleopteranand lepidoptera of the various sequences. Table code: “−” indicates noactivity seen; “+” indicates pesticidal activity; “NT” indicates nottested; “S” indicates stunt; “SS” indicates slight stunt; “LF” indicateslow feeding, “M” indicates mortality.

TABLE 3 Summary of Pesticidal Activity against Coleopteran andLepidoptera. APG Seq ID FAW CEW BCW ECB SWCB CPB Px WCR Mortality (%)APG00455 96 − − − − − NT NT 50-80% mortality APG00330 251 SS − − − − NTNT 0-50% mortality APG00359 68 − − − − − NT NT 0-50% mortality APG0034166 SS − − − − NT NT 0-50% mortality APG00224 28 SS − − − SS NT NT 0-50%mortality APG00562 126 − − − − − NT NT 0-50% mortality APG00410 79 SS −− − − NT NT 50-80% mortality APG00522 117 − − − − S NT NT 80-100%mortality APG00170 14 − − − NT NT NT NT 0-50% mortality APG00719 316 − −− − − NT NT 80-100% mortality APG00309 56 − − − − − NT NT 80-100%mortality APG00309 57 − − − − − NT NT 80-100% mortality APG00755 167 − −− − − NT NT 80-100% mortality APG00171 212 − − − − − NT NT 0-50%mortality APG00453 276 SS − − − − NT NT 50-80% mortality APG00420 267 SS− − − − NT NT 50-80% mortality APG00156 207 − − − − − NT NT 0-50%mortality APG00488 109 − − − − − NT NT 80-100% mortality APG00994 199 −− − − − NT NT 0-50% mortality APG00542 121 − − − − − NT NT 50-80%mortality APG00598 140 − − − − − NT NT 0-50% mortality APG00972 184 SS −− − − NT NT NT APG00343 5 SS − − − − NT NT NT APG00780 169 − − − NT NTNT NT 0-50% mortality APG00477 99 − − − NT NT NT NT 0-50% mortalityAPG00390 75 − − − NT NT NT NT 50-80% mortality APG00193 22 − SS − NT NTNT NT 50-80% mortality

Example 4 Pesticidal Activity Against Hemipteran

Protein Expression: Each of the sequences set forth in Table 4 wasexpressed in E. coli as described in Example 2. 400 mL of LB wasinoculated and grown to an OD600 of 0.6. The culture was induced with0.25 mM IPTG overnight at 16° C. The cells were spun down and the cellpellet was re-suspend in 5 mL of buffer. The resuspension was sonicatedfor 2 min on ice.

Second instar SGSB were obtained from a commercial insectary (BenzonResearch Inc., Carlisle, Pa.). A 50% v/v ratio of sonicated lysatesample to 20% sucrose was employed in the bioassay. Stretched parafilmwas used as a feeding membrane to expose the SGSB to the diet/samplemixture. The plates were incubated at 25° C.: 21° C., 16:8 day:nightcycle at 65% RH for 5 days.

Mortality was scored for each sample. The results are set forth in Table4. A dashed line indicates no mortality was detected. The proteinslisted in table 4 showed 25% mortality or 50% mortality (as indicated)against southern green stinkbug (1 stinkbug out of 4 died). The negativecontrols (empty vector expressed binding domain and buffer only) bothshowed no mortality (0 stinkbugs out of 4).

TABLE 4 Summary of Pesticidal Activity against Hemipteran APG Seq IDTested against SGSB APG00455 96 — APG00330 251 25% APG00359 68 —APG00341 66 — APG00224 28 — APG00562 126 — APG00410 79 — APG00522 117 —APG00170 14 — APG00719 316 — APG00309 56 — APG00309 57 25% APG00755 167— APG00171 212 — APG00453 276 — APG00420 267 — APG00156 207 — APG00488109 25% APG00994 199 25% APG00542 121 25% APG00598 140 — APG00972 184 —APG00343 5 — APG00780 169 — APG00477 99 — APG00390 75 — APG00193 22 —

Example 5 Transformation of Soybean

DNA constructs comprising each of SEQ ID NOs: 1-341 or active variantsor fragments thereof operably linked to a promoter active in a plant arecloned into transformation vectors and introduced into Agrobacterium asdescribed in PCT Application PCT/US2015/066702, filed Dec. 18, 2015,herein incorporated by reference in its entirety.

Four days prior to inoculation, several loops of Agrobacterium arestreaked to a fresh plate of YEP* medium supplemented with theappropriate antibiotics** (spectinomycin, chloramphenicol andkanamycin). Bacteria are grown for two days in the dark at 28° C. Aftertwo days, several loops of bacteria are transferred to 3 ml of YEPliquid medium with antibiotics in a 125 ml Erlenmeyer flask. Flasks areplaced on a rotary shaker at 250 RPM at 28° C. overnight. One day beforeinoculation, 2-3 ml of the overnight culture were transferred to 125 mlof YEP with antibiotics in a 500 ml Erlenmeyer flask. Flasks are placedon a rotary shaker at 250 RPM at 28° C. overnight.

Prior to inoculation, the OD of the bacterial culture is checked at OD620. An OD of 0.8-1.0 indicates that the culture is in log phase. Theculture is centrifuged at 4000 RPM for 10 minutes in Oakridge tubes. Thesupernatant is discarded and the pellet is re-suspended in a volume ofSoybean Infection Medium (SI) to achieve the desired OD. The culturesare held with periodic mixing until needed for inoculation.

Two or three days prior to inoculation, soybean seeds are surfacesterilized using chlorine gas. In a fume hood, a petri dish with seedsis placed in a bell jar with the lid off. 1.75 ml of 12 N HCl is slowlyadded to 100 ml of bleach in a 250 ml Erlenmeyer flask inside the belljar. The lid is immediately placed on top of the bell jar. Seeds areallowed to sterilize for 14-16 hours (overnight). The top is removedfrom the bell jar and the lid of the petri dish is replaced. The petridish with the surface sterilized is then opened in a laminar flow foraround 30 minutes to disperse any remaining chlorine gas.

Seeds are imbibed with either sterile DI water or soybean infectionmedium (SI) for 1-2 days. Twenty to 30 seeds are covered with liquid ina 100×25 mm petri dish and incubated in the dark at 24° C. Afterimbibition, non-germinating seeds are discarded.

Cotyledonary explants are processed on a sterile paper plate withsterile filter paper dampened using SI medium employing the methods ofU.S. Pat. No. 7,473,822, herein incorporated by reference.

Typically, 16-20 cotyledons are inoculated per treatment. The SI mediumused for holding the explants is discarded and replaced with 25 ml ofAgrobacterium culture (OD 620=0.8−20). After all explants are submerged,the inoculation is carried out for 30 minutes with periodic swirling ofthe dish. After 30 minutes, the Agrobacterium culture is removed.

Co-cultivation plates are prepared by overlaying one piece of sterilepaper onto Soybean Co-cultivation Medium (SCC). Without blotting, theinoculated cotyledons are cultured adaxial side down on the filterpaper. Around 20 explants can be cultured on each plate. The plates aresealed with Parafilm and cultured at 24° C. and around 120 μmoles m-2s-1(in a Percival incubator) for 4-5 days.

After co-cultivation, the cotyledons are washed 3 times in 25 ml ofSoybean Wash Medium with 200 mg/l of cefotaxime and timentin. Thecotyledons are blotted on sterile filter paper and then transferred toSoybean Shoot Induction Medium (SSI). The nodal end of the explant isdepressed slightly into the medium with distal end kept above thesurface at about 45 deg. No more than 10 explants are cultured on eachplate. The plates are wrapped with Micropore tape and cultured in thePercival at 24° C. and around 120 μmoles m-2s-1.

The explants are transferred to fresh SSI medium after 14 days. Emergingshoots from the shoot apex and cotyledonary node are discarded. Shootinduction is continued for another 14 days under the same conditions.

After 4 weeks of shoot induction, the cotyledon is separated from thenodal end and a parallel cut is made underneath the area of shootinduction (shoot pad). The area of the parallel cut is placed on SoybeanShoot Elongation Medium (SSE) and the explants cultured in the Percivalat 24° C. and around 120 μmoles m-2s-1. This step is repeated every twoweeks for up to 8 weeks as long as shoots continue to elongate.

When shoots reach a length of 2-3 cm, they are transferred to SoybeanRooting Medium (SR) in a Plantcon vessel and incubated under the sameconditions for 2 weeks or until roots reach a length of around 3-4 cm.After this, plants are transferred to soil.

Note, all media mentioned for soybean transformation are found in Paz etal. (2010) Agrobacterium-mediated transformation of soybean and recoveryof transgenic soybean plants; Plant Transformation Facility of IowaState University, which is herein incorporated by reference in itsentirety. (See, agron—www.agron.iastate.edu/ptf/protocol/Soybean.pdf.)

Example 6 Transformation of Maize

Maize ears are best collected 8-12 days after pollination. Embryos areisolated from the ears, and those embryos 0.8-1.5 mm in size arepreferred for use in transformation. Embryos are plated scutellumside-up on a suitable incubation media, such as DN62A5S media (3.98 g/LN6 Salts; 1 mL/L (of 1000× Stock) N6 Vitamins; 800 mg/L L-Asparagine;100 mg/L Myo-inositol; 1.4 g/L L-Proline; 100 mg/L Casamino acids; 50g/L sucrose; 1 mL/L (of 1 mg/mL Stock) 2,4-D). However, media and saltsother than DN62A5S are suitable and are known in the art. Embryos areincubated overnight at 25° C. in the dark. However, it is not necessaryper se to incubate the embryos overnight.

The resulting explants are transferred to mesh squares (30-40 perplate), transferred onto osmotic media for about 30-45 minutes, and thentransferred to a beaming plate (see, for example, PCT Publication No.WO/0138514 and U.S. Pat. No. 5,240,842). DNA constructs designed toexpress the GRG proteins of the present invention in plant cells areaccelerated into plant tissue using an aerosol beam accelerator, usingconditions essentially as described in PCT Publication No. WO/0138514.After beaming, embryos are incubated for about 30 min on osmotic media,and placed onto incubation media overnight at 25° C. in the dark. Toavoid unduly damaging beamed explants, they are incubated for at least24 hours prior to transfer to recovery media. Embryos are then spreadonto recovery period media, for about 5 days, 25° C. in the dark, andthen transferred to a selection media. Explants are incubated inselection media for up to eight weeks, depending on the nature andcharacteristics of the particular selection utilized. After theselection period, the resulting callus is transferred to embryomaturation media, until the formation of mature somatic embryos isobserved. The resulting mature somatic embryos are then placed under lowlight, and the process of regeneration is initiated by methods known inthe art. The resulting shoots are allowed to root on rooting media, andthe resulting plants are transferred to nursery pots and propagated astransgenic plants.

Example 7 Pesticidal Activity Against Nematodes

Heterodera Glycine's (Soybean Cyst Nematode) In Vitro Assay

Soybean Cyst Nematodes are dispensed into a 96 well assay plate with atotal volume of 100 uls and 100 J2 per well. The protein of interest asset forth in any one of SEQ ID NOs: 1-341 is dispensed into the wellsand held at room temperature for assessment. Finally, the 96 well platecontaining the SCN J2 is analyzed for motility. Data is reported as %inhibition as compared to the controls. Hits are defined as greater orequal to 70% inhibition.

Heterodera Glycine's (Soybean Cyst Nematode) on-plant Assay

Soybean plants expressing one or more of SEQ ID NOs: 1-341 are generatedas described elsewhere herein. A 3-week-old soybean cutting isinoculated with 5000 SCN eggs per plant. This infection is held for 70days and then harvested for counting of SCN cyst that has developed onthe plant. Data is reported as % inhibition as compared to the controls.Hits are defined as greater or equal to 90% inhibition.

Meloidogyne Incognita (Root-Knot Nematode) In Vitro Assay

Root-Knot Nematodes are dispensed into a 96 well assay plate with atotal volume of 100 uls and 100 J2 per well. The protein of interestcomprising any one of SEQ ID NOs: 1-341 is dispensed into the wells andheld at room temperature for assessment. Finally the 96 well platecontaining the RKN J2 is analyzed for motility. Data is reported as %inhibition as compared to the controls. Hits are defined as greater orequal to 70% inhibition.

Meloidogyne Incognita (Root-Knot Nematode) on-Plant Assay

Soybean plants expressing one or more of SEQ ID NOs: 1-341 are generatedas described elsewhere herein. A 3-week-old soybean is inoculated with5000 RKN eggs per plant. This infection is held for 70 days and thenharvested for counting of RKN eggs that have developed in the plant.Data is reported as % inhibition as compared to the controls. Hits aredefined as greater or equal to 90% inhibition.

Example 8 Additional Assays for Pesticidal Activity

The various polypeptides set forth in SEQ ID NOs: 1-341 can be tested toact as a pesticide upon a pest in a number of ways. One such method isto perform a feeding assay. In such a feeding assay, one exposes thepest to a sample containing either compounds to be tested or controlsamples. Often this is performed by placing the material to be tested,or a suitable dilution of such material, onto a material that the pestwill ingest, such as an artificial diet. The material to be tested maybe composed of a liquid, solid, or slurry. The material to be tested maybe placed upon the surface and then allowed to dry. Alternatively, thematerial to be tested may be mixed with a molten artificial diet, andthen dispensed into the assay chamber. The assay chamber may be, forexample, a cup, a dish, or a well of a microtiter plate.

Assays for sucking pests (for example aphids) may involve separating thetest material from the insect by a partition, ideally a portion that canbe pierced by the sucking mouth parts of the sucking insect, to allowingestion of the test material. Often the test material is mixed with afeeding stimulant, such as sucrose, to promote ingestion of the testcompound.

Other types of assays can include microinjection of the test materialinto the mouth, or gut of the pest, as well as development of transgenicplants, followed by test of the ability of the pest to feed upon thetransgenic plant. Plant testing may involve isolation of the plant partsnormally consumed, for example, small cages attached to a leaf, orisolation of entire plants in cages containing insects.

Other methods and approaches to assay pests are known in the art, andcan be found, for example in Robertson and Preisler, eds. (1992)Pesticide bioassays with arthropods, CRC, Boca Raton, Fla.Alternatively, assays are commonly described in the journals ArthropodManagement Tests and Journal of Economic Entomology or by discussionwith members of the Entomological Society of America (ESA). Any one ofSEQ ID NOS: 1-341 can be expressed and employed in an assay as set forthin Examples 3 and 4, herein.

All publications and patent applications mentioned in the specificationare indicative of the level of skill of those skilled in the art towhich this invention pertains. All publications and patent applicationsare herein incorporated by reference to the same extent as if eachindividual publication or patent application was specifically andindividually indicated to be incorporated by reference.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity ofunderstanding, it will be obvious that certain changes and modificationsmay be practiced within the scope of the appended claims.

That which is claimed is:
 1. A method for growing a plant in an areainfested with, or susceptible to infestation by, hemipteran insectpests, comprising: (a) planting a seed or a plant in an area infestedwith, or susceptible to infestation by, hemipteran insect pests, whereinthe seed or the plant, or a cell of either thereof, expresses anucleotide sequence that encodes an insecticidal polypeptide, whereinthe nucleotide sequence encodes a polypeptide comprising an amino acidsequence having at least 95% sequence identity to the amino acidsequence of SEQ ID NO:98, wherein said polypeptide is pesticidal againstat least one hemipteran pest; and (b) growing the seed or the plant. 2.The method of claim 1, wherein the nucleotide sequence encodes apolypeptide comprising the amino acid sequence of SEQ ID NO:98.
 3. Themethod of claim 1, wherein the seed or the plant is a monocot seed or amonocot plant.
 4. The method of claim 1, wherein the seed or the plantis a dicot seed or a dicot plant.
 5. The method of claim 1, wherein theplant is selected from the group consisting of corn (maize), sorghum,wheat, sunflower, tomato, crucifers, peppers, potato, cotton, rice,soybean, sugarbeet, sugarcane, tobacco, barley, and oilseed rape,Brassica sp., alfalfa, rye, millet, safflower, peanuts, sweet potato,cassava, coffee, coconut, pineapple, citrus trees, cocoa, tea, banana,avocado, fig, guava, mango, olive, papaya, cashew, macadamia, almond,oats, vegetables, ornamentals, and conifers.
 6. The method of claim 1,wherein the seed or the plant is a vegetable seed or vegetable plantselected from the group consisting of tomatoes, lettuce, green beans,lima beans, peas, and members of the genus Cucumis.
 7. The method ofclaim 1, wherein the seed or the plant is selected from the groupconsisting of azalea, hydrangea, hibiscus, roses, tulips, daffodils,petunias, carnation, poinsettia, and chrysanthemum.